| acidic ph induces fusion of cells infected with baculovirus to form syncytia. | the enveloped baculovirus insect cell system has been used extensively for expression of recombinant proteins, including viral fusion proteins. we tested wild-type baculovirus for endogenous fusion protein activity. syncytia formation, dye transfer, and capacitance changes were observed after incubating infected spodoptera frugiperda cells in acidic media, consistent with fusion protein activity. only a short acidic pulse of 10 s is needed to trigger syncytia formation. identical results were ob ... | 1992 | 1618326 |
| a baculovirus dual expression vector derived from the autographa californica nuclear polyhedrosis virus polyhedrin and p10 promoters: co-expression of two influenza virus genes in insect cells. | a baculovirus transfer vector, pacuw3, was developed to facilitate the insertion of two influenza virus genes, those encoding the haemagglutinin (ha) and neuraminidase (na) membrane glycoproteins, into the autographa californica nuclear polyhedrosis virus genome in a single cotransfection experiment. the na gene was inserted in place of the polyhedrin coding sequences under the control of the polyhedrin promoter, whereas the ha gene was placed under the control of a copy of the p10 promoter at a ... | 1991 | 1765769 |
| structural comparison of the autographa californica nuclear polyhedrosis virus-induced rna polymerase and the three nuclear rna polymerases from the host, spodoptera frugiperda. | a partial subunit structure has been determined for the novel rna polymerase that is induced in fall armyworm (spodoptera frugiperda) cells upon infection with the autographa californica nuclear polyhedrosis virus (acnpv). the putative structure includes nine polypeptides; the complexity of this structure is in accord with the high sedimentation coefficient (15s) estimated for this enzyme. a comparison of the putative structure of the virus-induced polymerase with those of the three host nuclear ... | 1991 | 1767583 |
| expression of hepatitis b virus surface antigen gene in insect baculovirus vector systems. | the gene coding for the hepatitis b virus surface antigen (hbsag) under the control of autographa californicanuclear polyhedrosis virus polyhedrin promoter was successfully inserted into the genome of the trichoplusia ni nuclear polyhdrosis virus. infection of spodoptera frugiperda cells with this recombinant virus produced a significant amount of hbsag protein and secreted 22 nm particles containing the hbsag. the expression of hbsag gene was also obtained both in trichoplusia ni larvae and in ... | 1991 | 1773016 |
| diagnostic potential of baculovirus-expressed rubella virus envelope proteins. | the envelope glycoproteins e1 and e2 of rubella virus were abundantly expressed in spodoptera frugiperda sf9 insect cells by using a baculovirus expression vector. the recombinant protein products were purified by immunoaffinity chromatography and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and enzyme immunoassay (eia). the purified recombinant antigen consisted of the envelope polypeptides, corresponding to the viral e1 and e2 proteins, and a poly ... | 1991 | 1774311 |
| foreign gene expression by a baculovirus vector with an expanded host range. | a nuclear polyhedrosis virus (npv) (baculoviridae)-based gene expression system was improved by dna recombination. the bmn cell line established from bombyx mori and the sf21 cell line established from spodoptera frugiperda are non-permissive for autographa californica multicapsid npv (acmnpv) and b. mori npv (bmnpv) replication, respectively. after cotransfection of acmnpv dna and bamhi-digested bmnpv dna into sf21 cells, progeny viruses were isolated by plaque purification on bmn cell monolaye ... | 1992 | 1629707 |
| expression of recombinant soluble and membrane-bound catechol o-methyltransferase in eukaryotic cells and identification of the respective enzymes in rat brain. | the rat and human recombinant soluble and membrane-bound catechol o-methyltransferase (s- and mb-comt, respectively) were expressed using mammalian and baculovirus vectors. low levels of rat and human s-comt polypeptides were detected by immunoprecipitation in k-562 cell lines transfected with the s-comt vectors. from k-562 cells transfected with the rat mb-comt construct, both s- and mb-comt recombinant proteins were detected by a rat comt-specific anti-serum. infection of lepidopteran spodopte ... | 1992 | 1633830 |
| baculovirus expression and characterization of catalytically active horseradish peroxidase. | studies of horseradish peroxidase (hrp), a prototypical enzyme, have provided much of the information that is available on the mechanisms and functions of hemoprotein peroxidases. hrp itself is widely used in biotechnological applications. further progress in defining the structure and function of the enzyme, however, requires its expression in a heterologous system. we report here baculovirus-mediated, high yield expression of a synthetic gene for hrp in spodoptera frugiperda cell culture. expr ... | 1992 | 1637184 |
| overproduction of rat 1,25-dihydroxyvitamin d3 receptor in insect cells using the baculovirus expression system. | the rat 1,25-dihydroxyvitamin d3 [1,25-(oh)2d3] receptor has been expressed at elevated levels in spodoptera frugiperda cells using the baculovirus expression vector system. the recombinant 1,25-(oh)2d3 receptor is full-length, binds 1,25-(oh)2d3, and is recognized by a monoclonal antibody specific for 1,25-(oh)2d3 receptor. densitometric scanning of coomassie brilliant blue-stained sds/polyacrylamide gels indicated a recombinant receptor protein level comprising 5% of the total soluble protein ... | 1991 | 1650474 |
| secretion of single-chain urokinase-type plasminogen activator from insect cells. | a cdna encoding human urokinase-type plasminogen activator was inserted downstream from the polyhedrin promoter of the baculovirus autographa californica nuclear polyhedrosis virus. a protein of similar mr to urokinase (uk) was synthesized and approx. 90% was secreted from recombinant virus-infected spodoptera frugiperda cells. zymography and western blotting analysis of the insect-derived protein demonstrated that it was comprised solely of the high-mr form of uk. no low-mr uk was detected. ami ... | 1991 | 1834526 |
| high-level synthesis and fate of acetylcholine in baculovirus-infected cells: characterization and purification of recombinant rat choline acetyltransferase. | rat choline acetyltransferase (chat) has been expressed at a high level in spodoptera frugiperda sf9 cells using a baculovirus expression system. a cdna containing the coding sequence for chat was inserted into the transfer vector pacym1 to yield the recombinant vector pacym1/chat. sf9 cells were then coinfected with pacym1/chat and the wild-type autographa californica virus. one recombinant virus particle, containing the cdna for chat, was selected that expressed a protein of 68.5 kda. forty ho ... | 1992 | 1548478 |
| differential factor binding at the promoter of early baculovirus gene pe38 during viral infection: gata motif is recognized by an insect protein. | regulatory elements interacting with dna-binding proteins have been investigated in the promoter sequence of the early pe38 gene in the autographa californica nuclear polyhedrosis virus (acnpv). a gata motif located 50 nucleotides upstream of the pe38 transcriptional start site is recognized differentially in the course of infection. as demonstrated by footprint and gel mobility shift assays, the gata sequences ttatct are protected by nuclear extracts from uninfected spodoptera frugiperda cells ... | 1992 | 1583720 |
| expression and characterization of recombinant mouse beta 2-microglobulin type a in insect cells infected with recombinant baculoviruses. | the murine beta 2-microglobulina cdna was cloned into pac373 and pvl941 transfer vectors and introduced via homologous recombination into the genome of autographa californica nuclear polyhedrosis virus downstream of the polyhedrin promoter. both types of recombinant baculoviruses were isolated and used to infect spodoptera frugiperda (sf9) lepidopteran cells. beta 2m was synthesized at a substantially higher rate in cells infected with the pvl941-derived virus than when the pac373-based virus wa ... | 1991 | 1754712 |
| expression of rotavirus proteins encoded by alternative open reading frames of genome segment 11. | the nucleotide sequence of rotavirus genome segment 11 shows that this gene contains three potential open reading frames. we used several approaches to determine whether any polypeptides other than ns26, the primary protein product, are expressed. in particular, we sought to determine whether the strong out-of-phase start codon present at nucleotides 80-82, which would encode a protein of 92 amino acids, is used in vivo or in cell-free systems. several modifications of gene 11 were made and foun ... | 1991 | 1847258 |
| high-level expression of the epstein-barr virus alkaline deoxyribonuclease using a recombinant baculovirus: application to the diagnosis of nasopharyngeal carcinoma. | the epstein-barr virus (ebv) alkaline deoxyribonuclease (dnase) was inserted into the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv). infection of the insect cell line spodoptera frugiperda (sf9) with the recombinant virus led to the expression of an enzymatically active alkaline dnase. the recombinant ebv alkaline dnase was highly soluble, and the recombinant baculovirus produced approximately 10-20 mg of ebv dnase per 1 x 10(9) cells. the recombinant enzyme activity was ... | 1991 | 1847261 |
| evidence for dissimilar properties of comoviral and picornaviral rna polymerases. | the poliovirus rna polymerase has been synthesized in spodoptera frugiperda cells by using the baculovirus expression system. crude sonicates of these cells exhibited an rna-elongating activity of a synthetic oligo(u) primer with poly(a) or cowpea mosaic virus (cpmv) rna as a template. a similar polymerase activity was found in extracts of insect cells in which foot-and-mouth disease virus (fmdv) proteins, including the putative polymerase, were produced. the analogous cpmv 87k protein and sever ... | 1991 | 1848591 |
| expression of the outer capsid protein vp5 of two bluetongue viruses, and synthesis of chimeric double-shelled virus-like particles using combinations of recombinant baculoviruses. | we have previously reported the assembly of virus-like particles (vlps), consisting of the four major structural proteins of bluetongue virus (btv), in spodoptera frugiperda cells coinfected with recombinant baculoviruses (french et al. (1990). j. virol. 64, 5695-5700). in this paper we report further studies using this system to assemble heterologous vlps containing the outer capsid proteins (vp2 and vp5) of a range of different btv serotypes. s. frugiperda cells were coinfected with three reco ... | 1991 | 1850928 |
| expression of rotavirus vp2 produces empty corelike particles. | the complete vp2 gene of bovine rotavirus strain rf has been inserted into the baculovirus transfer vector pvl941 under the control of the polyhedrin promoter. cotransfection of spodoptera frugiperda 9 cells with wild-type baculovirus dna and transfer vector dna led to the formation of recombinant baculoviruses which contain bovine rotavirus gene 2. infection of s. frugiperda cells with this recombinant virus resulted in the production of a protein similar in size and antigenic properties to the ... | 1991 | 1851866 |
| characterization and purification of human beta-galactosidase overexpressed in recombinant baculovirus-infected spodoptera frugiperda cells. | | 1991 | 1779628 |
| dengue-1 virus envelope glycoprotein gene expressed in recombinant baculovirus elicits virus-neutralizing antibody in mice and protects them from virus challenge. | in order to test the feasibility of baculovirus (autographa californica nuclear polyhedrosis virus, acnpv) expression vectors for making immunogens against dengue-1 (den-1) virus, a portion of the envelope (e) glycoprotein gene of den-1 virus was cloned and expressed. the recombinant baculovirus contains 107 nucleotides from the 3' terminus of the den-1 matrix (m) gene, which encodes a hydrophobic signal peptide and extends through the first 1, 245 nucleotides of e, terminating 243 nucleotides b ... | 1991 | 1877710 |
| cysteine-86 is not needed for the enzymic activity of glutathione s-transferase 3-3. | recombinant glutathione s-transferase 3-3 expressed in spodoptera frugiperda (sf9) cells with the use of a baculovirus expression system was modified with 1 mm-iodoacetamide. amino acid analysis indicated that 0.79 +/- 0.15 cysteine residue was modified per enzyme subunit. the s-carbaminomethylated protein retains the gsh-conjugating activity. glutathione s-transferase 3-3 modified with iodo[14c]acetamide was digested with achromobacter proteinase i and the resulting peptides were separated by h ... | 1991 | 1883338 |
| molecular cloning of a human basic fibroblast growth factor receptor cdna and expression of a biologically active extracellular domain in a baculovirus system. | a cdna clone encoding a human fibroblast growth factor (fgf) receptor was isolated from a hepatoma cell line cdna library. the cdna encodes a three immunoglobulinlike-domain fgf receptor that is similar to a human placental fgf receptor cdna but lacks two amino acids. the variation observed at these two amino acids, also seen in the two immunoglobulinlike-domain fgf-receptors, can be explained by an alternate splicing mechanism. we have used a baculovirus expression system to produce high levels ... | 1991 | 1662973 |
| lepidopteran cell variants resistant to 5-bromodeoxyuridine and their use for transfection of the hsv-tk gene. | following mutagenesis of cultured lepidopteran cells (spodoptera frugiperda) by ethylmethanesulfonate, three variants resistant to 5-bromodeoxyuridine (brdurd) were isolated. these clones were 100- to 200-fold more resistant to brdurd than the parental cells and were shown to be deficient in thymidine kinase (tk). the drug-resistant phenotype was stable for up to three years of culture under nonselective conditions. it was also found that the s. frugiperda cell line was highly resistant to amino ... | 1991 | 1665497 |
| functional expression of the human growth factor activatable na+/h+ antiporter (nhe-1) in baculovirus-infected cells. | we constructed a recombinant baculovirus, based on autographa californica nuclear polyhedrosis virus, containing the human na+/h+ antiporter cdna under control of the polyhedrin promoter. when infected with this recombinant baculovirus, the sf9 cell line, derived from spodoptera frugiperda, expresses a fully functional na+/h+ antiporter as measured by the generation of an amiloride-sensitive na+ influx in response to an acid load. the na+/h(+)-exchange activity, not detectable in sf9 cells, emer ... | 1991 | 1654101 |
| overexpression and characterization of the human mineralocorticoid receptor. | the full-length human renal mineralocorticoid receptor (hmr) has been overproduced in spodoptera frugiperda (sf9) insect cells using baculovirus-mediated expression. the overproduced hmr binds aldosterone with high affinity (kd = 1.36 nm) and has high affinity for cortisol, cortexolone, and progesterone. immunoprecipitation and immunoblot analysis of the recombinant hmr with mr-specific antibodies reveal three major protein bands with molecular masses of 115, 119, and 125 kda. hmr isoforms show ... | 1991 | 1655735 |
| baculovirus-mediated expression of the human vitamin d receptor. functional characterization, vitamin d response element interactions, and evidence for a receptor auxiliary factor. | a baculovirus expression vector system (bevs) was used to overproduce the full-length human vitamin d receptor (hvdr) in spodoptera frugiperda ovarian cells. hvdr was expressed to a level of 0.5% of the total soluble protein in this system. western analysis demonstrated that the baculovirus-generated protein had electrophoretic and immunologic properties equivalent to those of hvdr expressed in mammalian cells. the bevs-derived receptor displayed specificity and high affinity (apparent kd = 0.7 ... | 1991 | 1655763 |
| characterization of an active, non-myristylated, cytoplasmic form of the lymphoid protein tyrosine kinase pp56lck. | pp56lck is a member of the src family of tyrosine kinases mainly expressed in t lymphocytes. src tyrosine kinases have been implicated in the control of cell growth and differentiation in different cell types, but the mechanism of regulation of these enzymes is poorly understood. in order to characterize the distinct species of pp56lck, we have produced high yields of enzymatically active wild type pp56lck using the eukaryotic baculovirus expression system in spodoptera frugiperda insect cells ( ... | 1991 | 1911540 |
| dna restriction polymorphism in wild isolates of spodoptera frugiperda nuclear polyhedrosis virus. | restriction endonuclease analysis was used to examine variation in dna of 22 wild isolates of spodoptera frugiperda nuclear polyhedrosis virus (sfnpv). eleven of the 15 isolated from louisiana were distinguishable based on restriction fragment profiles from the enzymes bamhi, hindiii, and ecori. there was significant genetic variation in sfnpv isolates within single agricultural fields. nucleotide sequence divergence values, based on restriction fragment profiles, indicated that genetic variatio ... | 1991 | 1679452 |
| characterization of pseudorabies virus neutralization antigen glycoprotein giii produced in insect cells by a baculovirus expression vector. | the gene encoding the complete glycoprotein of pseudorabies virus (prv, yamagata-s81 strain glycoprotein giii) has been inserted into the baculovirus transfer vector pacym1s derived from the nuclear polyhedrosis virus of autographa californica (acnpv). a spodoptera frugiperda cell line, sf21ae, was efficiently co-transfected with the transfer vector containing the giii gene and acnpv dna by cationic liposomes (lipofectin). the gene was placed under the control of the acnpv polyhedrin promoter an ... | 1991 | 1661981 |
| categorizing some early and late transcripts directed by the autographa californica nuclear polyhedrosis virus. | using an s1 mapping assay on rna from spodoptera frugiperda cells infected by the autographa californica nuclear polyhedrosis virus in the presence and absence of cycloheximide and aphidicolin, we can distinguish three classes of transcripts. first, there are those whose synthesis is blocked by the dna synthesis inhibitor aphidicolin and which are therefore late transcripts. these include the late transcript of the 39k gene and a late leftward transcript across the xhoi site in the hindiii-f reg ... | 1990 | 1698929 |
| characterization of yellow fever virus proteins e and ns1 expressed in vero and spodoptera frugiperda cells. | the cdna encoding the e and ns1 proteins of the yellow fever virus (yfv) was expressed in spodoptera frugiperda cells via the recombinant baculovirus ac-e. ns1 as a gp100 precursor which was cleaved to generate the recombinant proteins e and ns1 similar in size, folding and antigenicity to the authentic ones. recombinant protein e exhibited immunodominant epitopes as judged by its reactivity with yfv-neutralizing mabs. using the triton x-114 phase separation system, authentic and recombinant e p ... | 1991 | 1710649 |
| enzyme activities in four different forms of human immunodeficiency virus 1 pol gene products. | five cassettes of the pol gene of human immunodeficiency virus 1 were constructed and inserted under the control of the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus by homologous recombination. the first cassette polf contains the full-length pol open reading frame; the second cassette pol100 starts with the first aug codon of the pol gene and deletes 103 amino acids from the amino terminus of the pol gene product; the third cassette pol97 deletes the entire prot ... | 1991 | 1711203 |
| characterization of haemagglutinin-neuraminidase glycoprotein of newcastle disease virus expressed by a recombinant baculovirus. | a recombinant baculovirus containing a cdna which encodes haemagglutinin-neuraminidase (hn) of newcastle disease virus (ndv) was constructed. spodoptera frugiperda cells infected with this recombinant virus produced a large amount of hn glycoprotein similar to the authentic hn in size. the recombinant hn glycoprotein was localized on the surface of the infected cells and conserved its haemadsorption and neuraminidase activities. the antigenic properties of the recombinant hn glycoprotein seemed ... | 1991 | 1927050 |
| molecular basis of thyroid hormone regulation of myelin basic protein gene expression in rodent brain. | regulation of myelin basic protein (mbp) gene expression by thyroid hormone has been investigated in rodent brain. quantitation of the 4 major alternatively spliced transcripts by rnase protection assay showed that the individual mrnas, corresponding to mbp isoforms 21.5, 18.5, 17, and 14 kda, were decreased from 2- to 17-fold at all ages studied (4-60 days) in hypothyroid animals when compared to euthyroid, but the timing of onset of expression was not altered. mbp mrna was also reduced in youn ... | 1991 | 1720778 |
| expression and purification of functional g protein alpha subunits using a baculovirus expression system. | the alpha subunits of the heterotrimeric guanine nucleotide-binding proteins gi1, gi2, gi3, g0, and gs have been overexpressed in sf9 cells using a baculovirus expression system. the gi1 alpha, gi2 alpha, gi3 alpha, and g0 alpha have been purified to homogeneity from infected spodoptera frugiperda (sf9) cells and characterized. yields of up to 1.8 mg of purified recombinant g alpha have been obtained from 300-ml cultures of infected cells. the recombinant alpha subunits are myristoylated and are ... | 1992 | 1730649 |
| synthesis and toxicity of full-length and truncated bacterial cryivd mosquitocidal proteins expressed in lepidopteran cells using a baculovirus vector. | full-length (72k) and truncated (61k) cryivd mosquitocidal proteins of bacillus thuringiensis (bt) were expressed in spodoptera frugiperda cells and larvae of trichoplusia ni using a baculovirus vector to investigate the role of cryivd peptides in toxicity as well as to evaluate further the baculovirus/lepidopteran system for expressing bt proteins. the cryivd genes were inserted into the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) under control of the polyhedrin ... | 1992 | 1730944 |
| identification of the human platelet gtpase activating protein for the cdc42hs protein. | the cdc42hs protein appears to be an isoform of the ras-related gtp-binding protein g25k and is an apparent human homolog of the saccharomyces cerevisiae cell-division-cycle protein, cdc42sc. in this study, we report the identification of a gtpase-activating protein (gap) for cdc42hs from human platelets (designated from here on as cdc42hs-gap). the cdc42hs-gap activity was solubilized from platelet membranes, recovered through successive chromatography steps (the final step being mono-q chromat ... | 1991 | 1939135 |
| expression of the mouse c-abl type iv proto-oncogene product in the insect cell baculovirus system. | the cellular gene c-abl is the normal homologue of the transforming gene (v-abl) within the genome of the abelson leukaemia virus. the cdna sequence coding for the cellular form of the murine abl gene (c-abl type iv) has been inserted into the baculovirus transfer vector, pac36c, so that the c-abl gene is under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). spodoptera frugiperda cells infected with the recombinant transfer vector in the prese ... | 1992 | 1737071 |
| baculovirus recombinant expressing a secreted form of a transmembrane carcinoma-associated antigen. | ga733-2 is a monoclonal antibody-defined, 40-kda glycoprotein antigen that is associated with carcinomas of various origins. hydrophobicity analysis of the protein sequence predicted by complementary dna (cdna) has suggested that the ga733-2 antigen is a type i membrane protein. in this study, the polymerase chain reaction was used in a strategy to omit cdna sequences for the transmembrane and cytoplasmic domains, thereby converting the extracellular domain into a secretory protein. full-length ... | 1992 | 1737342 |
| expression of the saccharomyces cerevisiae kex2p endoprotease in inset cells. evidence for a carboxy-terminal autoprocessing event. | the pheromone-processing kex2p endoprotease of saccharomyces cerevisiae has been difficult to characterize due to its low level of expression in yeast cells. to overcome this problem, we have overexpressed kex2p using the baculovirus/insect cell expression system. spodoptera frugiperda sf9 insect cells infected with a recombinant baculovirus, containing the complete kex2 gene which encodes the kex2p protease (814 amino acids), accumulate an 120-kda functional form of the enzyme. the inhibition p ... | 1992 | 1740121 |
| identification of putative insect brush border membrane-binding molecules specific to bacillus thuringiensis delta-endotoxin by protein blot analysis. | binding sites for insecticidal toxins of bacillus thuringiensis are located in the brush border membranes of insect midguts. two approaches were used to investigate the interactions of b. thuringiensis subsp. kurstaki hd-73 cryia(c) toxin with brush border membrane vesicles from sensitive and naturally resistant insects: 125i-toxin-vesicle binding assays and protein blots probed with 125i-cryia(c) toxin. in bioassays, manduca sexta and heliothis virescens larvae were highly sensitive, helicoverp ... | 1991 | 1746942 |
| expression of biologically active human corticosteroid binding globulin by insect cells: acquisition of function requires glycosylation and transport. | human corticosteroid binding globulin (hcbg) is a 50- to 55-kda serum glycoprotein that binds cortisol and progesterone with high affinity. to map the steroid-binding domain and to investigate the folding pathways of hcbg, we have established an expression system based on infection of insect cells with a recombinant baculovirus encoding hcbg. infected spodoptera frugiperda (sf9) cells secrete immunoreactive hcbg at high levels (16-24 pmol per 10(6) cells per 40 h), and the recombinant protein bi ... | 1991 | 1862072 |
| expression of rat tyrosine hydroxylase in insect tissue culture cells and purification and characterization of the cloned enzyme. | rat tyrosine hydroxylase has been expressed at high levels in spodoptera frugiperda cells using a baculovirus expression system. a cdna containing the coding region for pc12 tyrosine hydroxylase was inserted into the unique ecori site of the transfer vector pljc8 to yield the recombinant vector pljc9. spodoptera frugiperda cells were then co-infected with pljc9 and wild type autographa californica nuclear polyhedrosis virus. recombinant virus particles containing the cdna for tyrosine hydroxylas ... | 1990 | 1967606 |
| biological function of recombinant il-6 expressed in a baculovirus system. | the cdna of human interleukin-6 (il-6) was cloned into baculovirus dna. insect cells (spodoptera frugiperda cells) infected with the recombinant baculovirus secreted a large amount of 22k protein into the culture medium. this culture fluid contained high biological activity of growth stimulation of a mouse myeloid cell line (mh-60). the il-6 was purified by a one-step procedure employing immunoaffinity chromatography of monoclonal antibody to il-6. the specific activities (bsf-2 reference units/ ... | 1991 | 1883915 |
| retarded processing of influenza virus hemagglutinin in insect cells. | when expressed in spodoptera frugiperda cells by a baculovirus vector, the hemagglutinin of fowl plague virus has been found to contain palmitic acid in covalent hydroxylamine-sensitive linkage, indicating that these cells have the capacity to acylate foreign proteins at cysteine residues. centrifugation on sucrose density gradients and immune precipitation with conformation-specific antibodies were used to compare trimerization of the hemagglutinin in insect cells and in fowl plague virus-infec ... | 1991 | 1984645 |
| the s2 subunit of the spike glycoprotein of bovine coronavirus mediates membrane fusion in insect cells. | the hemagglutinin/esterase (he), spike precursor (s) and the s1 and s2 subunits of the spike precursor protein of bovine coronavirus were expressed in spodoptera frugiperda (sf9) cells, and the cell-fusing activity of each recombinant glycoprotein was examined. extensive syncytia formation was observed in cells infected with the s2 recombinant but not with the he or s1 recombinant baculoviruses. fusion of sf9 cells expressing the intact s protein precursor was evident after trypsin treatment. th ... | 1991 | 1984658 |
| processing, secretion, and immunoreactivity of carboxy terminally truncated dengue-2 virus envelope proteins expressed in insect cells by recombinant baculoviruses. | two recombinant baculoviruses were constructed by inserting via the transfer vector pacym1 the genes coding for the structural proteins of dengue (den)-2 virus downstream from the polyhedrin promoter of autographa californica nuclear polyhedrosis virus. the two recombinants differed in truncation of 26 and 71 amino acids, respectively, in the carboxy-terminal sequence of den-specific envelope (e) glycoprotein. recombinant den-2 e glycoproteins were processed and transported to the surface of spo ... | 1991 | 1984665 |
| characterization and functional properties of the a and b forms of human progesterone receptors synthesized in a baculovirus system. | human progesterone receptors (pr) were overexpressed in spodoptera frugiperda (sf9) insect cells using a recombinant baculovirus system. recombinant viruses were constructed that produced either full-length a (94k) or b (120k) forms of human pr, and each was expressed as a functional protein. steroid and dna binding activities were found to be indistinguishable from that of endogenous human pr in t47d breast cancer cells. moreover, as analyzed by gel-mobility shift, recombinant pr-a and pr-b eac ... | 1991 | 1779977 |
| identification of the very early transcribed baculovirus gene pe-38. | we have started to identify early viral rnas that are transcribed at 1 h after inoculation to investigate the mechanism involved in the regulation of early gene expression of autographa californica nuclear polyhedrosis virus (acnpv). cloned viral dna fragments were hybridized to northern (rna) blots of polyadenylated rna isolated from spodoptera frugiperda cells at 1, 2, and 6 h postinfection to localize very early transcripts. subsequently we prepared a cdna library of polyadenylated rna transc ... | 1991 | 1987375 |
| characterization of a recombinant single-chain molecule comprising the variable domains of a monoclonal antibody specific for human fibrin fragment d-dimer. | a recombinant single-chain molecule, scfv-k12g0, containing the variable domains of the monoclonal antibody ma-15c5, specific for fragment d-dimer of human cross-linked fibrin, was constructed and expressed in spodoptera frugiperda, sf9, insect cells. the arg108 carboxyl-terminal amino acid of the variable domain of the light-chain of the antibody was connected through a synthetic ala-gly-gln-gly-ser-ser-val peptide linker with the gln1 amino-terminal amino acid of the variable domain of its hea ... | 1991 | 1885569 |
| immunolocalization of alzheimer beta-amyloid peptide precursor to cellular membranes in baculovirus expression system. | one characteristic of alzheimer's disease (a beta disease) is the accumulation of amyloid deposits within the extracellular space of the brain and meninges. a 40 amino acid peptide called beta-peptide or a4 protein is the subunit of the amyloid fibrils found in these deposits. the sequence of beta-peptide is contained within those of a family of larger proteins called the alzheimer beta-amyloid peptide precursor (app). these apps contain, in addition to a signal sequence, a hydrophobic sequence ... | 1991 | 1787542 |
| purification and biochemical characterization of a soluble mouse interferon-gamma receptor produced in insect cells. | the extracellular domain of the mouse interferon gamma receptor comprising amino acids 17-243 of the protein was produced in spodoptera frugiperda cells infected with a recombinant baculovirus. the receptor was mainly secreted into the culture medium and was purified to homogeneity in several hundred milligram amounts. the purification procedure involved four chromatography steps and delivered a soluble and active receptor with an overall recovery of 30%. from each purification run, two pools of ... | 1991 | 1828230 |
| recombinant baculoviruses expressing yellow fever virus e and ns1 proteins elicit protective immunity in mice. | recently, we showed that yellow fever virus (yfv) e and ns1 proteins in spodoptera frugiperda cells infected with a recombinant baculovirus are similar, if not identical to those produced during yfv infection. to study the role of e and ns1 in the induction of protective immunity against fatal yfv challenge, these viral antigens were expressed either alone or in tandem via recombinant baculoviruses ac-e. ns1, ac-e1 and ac-ns1. swiss mice were immunized with lysates of insect cells infected with ... | 1991 | 1834798 |
| rabies virus nucleoprotein expressed in and purified from insect cells is efficacious as a vaccine. | a cdna copy of the rna gene that encodes the nucleoprotein n of rabies virus evelyn-rokitnicki-abelseth strain was cloned into baculovirus. the recombinant baculovirus expressed the n protein abundantly in spodoptera frugiperda cells. the n protein was extracted from infected spodoptera frugiperda cells and purified to near homogeneity by affinity chromatography. the purified n protein reacted with 31 of 32 monoclonal antibodies that recognize native rabies virus ribonucleoprotein. like the ribo ... | 1991 | 2000406 |
| chemical modification of bacillus thuringiensis subsp. thuringiensis (hd-524) trypsin-activated endotoxin: implication of tyrosine residues in lepidopteran cell lysis. | a purified protein fraction from a solubilized and trypsin-digested extract of bacillus thuringiensis subsp. thuringiensis (hd-524) fermentation powder was lytic to cells from several lepidopteran lines. maximum yield was obtained by alkaline carbonate-thiocyanate solubilization of washed powder followed by trypsin digestion and sephacryl (s-300) chromatography. the alkaline carbonate-solubilized fraction consisted predominantly of two bands on sodium dodecyl sulfate-polyacrylamide gel electroph ... | 1991 | 2002242 |
| synthesis of newcastle disease virus (ndv)-like envelopes in insect cells infected with a recombinant baculovirus expressing the haemagglutinin-neuraminidase of ndv. | electron microscopical examination of negatively stained extracellular fluids (ecf) from spodoptera frugiperda cell cultures infected with a recombinant baculovirus expressing the newcastle disease virus (ndv) haemagglutinin-neuraminidase (hn) revealed ndv-like envelopes which resembled the envelopes of authentic ndv. immunogold staining with anti-ndv hn monoclonal antibodies demonstrated hn antigen in spikes on the ndv-like envelopes. the ecf from the recombinant-infected cultures also containe ... | 1991 | 2005440 |
| dithyreanitrile: an unusual insect antifeedant from dithyrea wislizenii. | dithyreanitrile, a novel sulfur-containing indole alkaloid, was isolated from the seeds of dithyrea wislizenii (cruciferae). dithyreanitrile inhibits feeding of fall armyworm (spodoptera frugiperda) and european corn borer (ostrinia nubilalis) larvae. dithyreanitrile, the first natural product with two sulfur atoms and a nitrile attached to the same carbon, was characterized by x-ray diffraction, spectroscopy, and chemical synthesis. | 1991 | 2009944 |
| baculovirus-directed high level expression of the hepatitis delta antigen in spodoptera frugiperda cells. | the hepatitis delta antigen (hdag) is a multifunctional protein. it forms the core-like structure of the hepatitis delta virus (hdv) but also enhances replication of hdv in the nucleus of the hepatocyte. a cdna fragment encoding hdag was inserted adjacent to the polyhedrin promoter of autographa californica nuclear polyhedrosis virus present in the baculovirus transfer vector pvl941. after transfection of spodoptera frugiperda (sf9) cells a recombinant baculovirus ac delta 1 was isolated and pur ... | 1991 | 2016595 |
| expression and characterization of a rabbit liver cytochrome p450 belonging to p450iib subfamily with the aid of the baculovirus expression vector system. | using baculovirus a cdna for cytochrome p450 (p450f1) belonging to rabbit p450iib subfamily was expressed in spodoptera frugiperda cells, where p450f1 was located on electron-dense structures (derived from the endoplasmic reticulum) present in both the cytoplasm and nucleus. partially purified p450f1 exhibited absorption spectra similar to those of p450(1), the major phenobarbital-inducible form in rabbit liver. like p450(1), p450f1 could oxidize aminopyrine, benzphetamine, 7-ethoxycoumarin, 1-n ... | 1991 | 2018490 |
| synthesis of the virus-specified tubules of epizootic haemorrhagic disease virus using a baculovirus expression system. | the formation of virus-specific tubules is one of the most characteristic features in the orbivirus infection cycle, yet little is known about their role in virus replication. the tubuli are composed of a major nonstructural protein, ns1. we have investigated the expression of the ns1-encoding gene of epizootic haemorrhagic disease virus serotype 2 (alberta-strain) by producing a recombinant autographa californica nuclear polyhedrosis virus (acnpv). prior to cloning in the baculovirus transfer v ... | 1991 | 1891959 |
| purification and characterization of the recombinant human aldose reductase expressed in baculovirus system. | large quantities of recombinant human aldose reductase were produced using spodoptera frugiperda cells and properties of the enzyme were characterized. direct purification of the recombinant aldose reductase by affinity column chromatography using matrex gel orange a yielded a single 36 kda band, similar in size to the purified human muscle aldose reductase, on a sodium dodecyl sulfate-polyacrylamide gel after silver staining. the isoelectric point of the recombinant enzyme was 5.85 which is ide ... | 1991 | 1905957 |
| purification and characterization of human immunodeficiency virus type 1 nef gene product expressed by a recombinant baculovirus. | we have constructed the recombinant baculovirus which expresses the human immunodeficiency virus type 1 negative factor (nef) gene. spodoptera frugiperda cells infected with the recombinant virus produced a 27-kda protein which reacted with rabbit antisera raised against a carboxy-terminal synthetic peptide of the nef protein by immunoblot analysis. labeling experiment showed that the recombinant nef protein was myristoylated. the recombinant nef protein was purified to near homogeneity by deae- ... | 1991 | 1909480 |
| alpha-mannosidase-catalyzed trimming of high-mannose glycans in noninfected and baculovirus-infected spodoptera frugiperda cells (iplb-sf-21ae). a possible contributing regulatory mechanism for assembly of complex-type oligosaccharides in infected cells. | incubation of a spodoptera frugiperda (iplb-sf-21ae) cell extract with the oligosaccharide man9glcnac2, the aglucosyl derivative of the glycan that is normally transferred from the dolichol carrier to the relevant asn residue in the nascent protein, results in its trimming to man6glcnac2, an intermediate that is relatively stable to further alpha-d-mannosidase action in these cells. on the other hand, incubation of a similar extract from cells that had been infected for various times with a wild ... | 1991 | 1911772 |
| characterization of a chimeric plasminogen activator consisting of a single-chain fv fragment derived from a fibrin fragment d-dimer-specific antibody and a truncated single-chain urokinase. | an mr 57,000 single-chain chimeric plasminogen activator, k12g0s32, consisting of a variable region fragment (fv) derived from the fibrin fragment d-dimer-specific monoclonal antibody ma-15c5 and of a 33-kda (amino acids ala132 to leu411) recombinant single-chain urokinase-type plasminogen activator (rscu-pa-33k) was studied. k12g0s32, secreted by infected spodoptera frugiperda insect cells at a rate of 1.5 micrograms/10(6) cells/48 h, was purified to homogeneity by ion-exchange chromatography a ... | 1991 | 1918077 |
| transcriptional analyses of baculovirus polyhedrin and foreign gene expression relative to baculovirus p10 mrna levels. | comparisons have been made between the p10 and polyhedrin mrna levels recovered from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acnpv). in molar terms and from 18 h post-infection (p.i.), the polyhedrin mrna species increased to levels one and a half times to twice as high as the p10 levels. the influence of the polyhedrin leader sequence on the expression of a foreign gene under the control of the polyhedrin promoter was investigated using a ser ... | 1991 | 1919531 |
| the solubility of inclusion proteins from bacillus thuringiensis is dependent upon protoxin composition and is a factor in toxicity to insects. | bacillus thuringiensis subsp. aizawai hd133 is one of several strains particularly effective against plodia interpunctella selected for resistance to b. thuringiensis subsp. kurstaki hd1 (dipel). b. thuringiensis subsp. aizawai hd133 produces inclusions containing three protoxins, cryia(b), cryic, and cryid, and the cryic protoxin has been shown to be active on resistant p. interpunctella as well as on spodoptera larvae. the cryia(b) protoxin is very similar to the major one in b. thuringiensis ... | 1991 | 2059054 |
| baculovirus gp64 gene expression: analysis of sequences modulating early transcription and transactivation by ie1. | expression of the baculovirus major envelope glycoprotein gene (gp64) is regulated by transcription from both early and late promoters. to characterize the early promoter and identify sequences involved in the regulation of gp64 early transcription, promoter-reporter gene fusions were generated from the orygia pseudotsugata nuclear polyhedrosis virus gp64 promoter and were analyzed by transient expression in uninfected insect cells. for these analyses, 5' deletion mutations were constructed in t ... | 1991 | 1920618 |
| overexpression and purification of transcriptionally competent creb from a recombinant baculovirus. | signal transduction and viral stimulatory pathways converge ultimately at the level of transcriptional activation to influence the expression of a variety of cellular genes in response to environmental stimuli and developmental signals. recent studies have implicated the cyclic amp-responsive element-binding protein (creb) to be involved in mediating transcriptional activation in response to multiple varied stimuli, including (1) stimulation of the protein kinase a signal transduction pathway; ( ... | 1991 | 1840397 |
| expression of bovine herpesvirus 1 glycoprotein giv by recombinant baculovirus and analysis of its immunogenic properties. | the gene encoding the giv glycoprotein of bovine herpesvirus 1 has been inserted into the genome of autographa californica baculovirus in lieu of the coding region of the a. californica baculovirus polyhedrin gene. recombinant protein was identified by its reactivity with giv-specific monoclonal antibodies and expressed at high levels (about 85 micrograms per 2.5 x 10(6) cells) in spodoptera frugiperda (sf9) cells. the recombinant glycoprotein had an apparent molecular mass of 63 kda, indicating ... | 1991 | 1845887 |
| high-level expression of a mitochondrial enzyme, ornithine transcarbamylase from rat liver, in a baculovirus expression system. | the mitochondrial enzyme, ornithine transcarbamylase (otc) from rat liver was expressed in spodoptera frugiperda (sf) insect cells using a baculovirus vector. when insect cells were infected with recombinant autographica californica nuclear polyhedrosis virus (acnpv) containing a cdna encoding the precursor form of otc (potc) inserted into the polyhedrin gene, they expressed catalytically active enzyme at levels of approximately 2.5 micrograms/10(6) cells. about 25% of the active enzyme was a no ... | 1991 | 2069719 |
| expression of the human multidrug transporter in insect cells by a recombinant baculovirus. | the plasma membrane associated human multidrug resistance (mdr1) gene product, known as the 170-kda p-glycoprotein or the multidrug transporter, acts as an atp-dependent efflux pump for various cytotoxic agents. we expressed recombinant human multidrug transporter in a baculovirus expression system to obtain large quantities and further investigate its structure and mechanism of action. mdr1 cdna was inserted into the genome of the autographa californica nuclear polyhedrosis virus under the cont ... | 1990 | 1970935 |
| characterization of baculovirus-expressed rift valley fever virus glycoproteins synthesized in insect cells. | a cdna corresponding to the complete coding region of the m rna of the m12 mutant of rift valley fever virus (rvfv) strain zh548 (k. takehara, m-k. min, j.k. battles, k. sugiyama, v.c. emery, j.m. dalrymple, and d.h.l. bishop, virology, 169, 452-457, 1989) has been inserted into the baculovirus transfer vector pacym1. by comparison with the parent rvfv, the m rna of the m12 mutant has a new small open reading frame (orf1) upstream of the one that initiates the precursor of the viral glycoprotein ... | 1990 | 2077783 |
| proteins encoded in the 81.2- to 85.0-map-unit fragment of autographa californica nuclear polyhedrosis virus dna can be translated in vitro and in spodoptera frugiperda cells. | we have previously demonstrated that five open reading frames exist in the nucleotide sequence of the 81.2- to 85.0-map-unit (m.u.) segment of plaque isolate e of autographa californica nuclear polyhedrosis virus (acnpv) dna. the corresponding polypeptides are 9.8, 12.1, 36.6, 25.0, and 48.2 kda in size (c. oellig, b. happ, t. müller, and w. doerfler, j. virol. 63:1494, 1989), and we have investigated whether these proteins can be translated in infected cells. on subfragments of this viral dna s ... | 1991 | 1985220 |
| expression of the bacillus anthracis protective antigen gene by baculovirus and vaccinia virus recombinants. | the gene encoding bacillus anthracis protective antigen (pa) was modified by site-directed mutagenesis, subcloned into baculovirus and vaccinia virus plasmid transfer vectors (pacym1 and psc-11, respectively), and inserted via homologous recombinations into baculovirus autographa californica nuclear polyhedrosis virus or vaccinia virus (strains wr and connaught). expression of pa was detected in both systems by immunofluorescence assays with antisera from rabbits immunized with b. anthracis pa. ... | 1990 | 2105271 |
| response of nuclear polyhedrosis virus-resistant spodoptera frugiperda larvae to other pathogens and to chemical insecticides. | selection in the laboratory for spodoptera frugiperda (sf) resistant to nuclear polyhedrosis virus (npv) affected the susceptibility of the insect to certain other mortality agents, including a chemical insecticide. median lethal concentrations (lc50s) and associated statistics were compared for several mortality agents between colonies of npv-resistant and -susceptible (control) insects. compared to the susceptible insects, the npv-resistant insects were cross-resistant to the s. frugiperda gra ... | 1990 | 2108223 |
| identification and characterization of the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. | the ie-1 gene of orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) was mapped between 95.7 and 97.1 map units on the viral genome. sequence analysis of the opmnpv ie-1 gene (opie-1) identified an open reading frame that coded for a predicted protein of 560 amino acids with a molecular weight of 64,775. transcriptional analysis of opmnpv-infected lymantria dispar (ld652y) cells identified two rnas homologous to the opie-1 open reading frame that were 1.7 and 1.9 kb in size. the ... | 1991 | 1989381 |
| single-step purification and biological activity of human nerve growth factor produced from insect cells. | human nerve growth factor (ngf) was cloned and engineered for expression in a baculovirus-infected spodoptera frugiperda (sf-9) insect cell system. culture supernatants contained 2-3 mg/l of recombinant human ngf. the human ngf produced by this system was purified to apparent homogeneity with a single-step affinity chromatography procedure using a high-affinity monoclonal antibody originally raised against murine ngf. the purification procedure yielded 1-2 mg of pure, human ngf per liter of cult ... | 1991 | 1993887 |
| expression of biologically active human antithrombin iii by recombinant baculovirus in spodoptera frugiperda cells. | antithrombin iii (atiii) is a plasma-borne serine protease inhibitor that plays a pivotal role in the regulation of hemostasis. the cdna for atiii has been available, but genetic studies on the functional domains of atiii have not progressed because of the absence of an expression system that will yield sufficient quantities of biologically active protein for biochemical analyses. in the present studies the cdna of the human antithrombin iii gene was inserted into the vector pvl 1393, which is s ... | 1991 | 1995647 |
| sequential rearrangement and nuclear polymerization of actin in baculovirus-infected spodoptera frugiperda cells. | proper assembly of nucleocapsids of the baculovirus autographa californica nuclear polyhedrosis virus is prevented by cytochalasin d, a drug that interferes with actin microfilament function. to investigate the involvement of microfilaments in a. californica nuclear polyhedrosis virus replication, a fluorescence microscopy study was conducted that correlated changes in distribution of microfilaments with events in the life cycle of the virus. tetramethylrhodamine isothiocyanate-labeled phalloidi ... | 1991 | 1995943 |
| expression and characterization of a functional human immunodeficiency virus envelope glycoprotein in insect cells. | recombinant baculoviruses were used to express the gp160 envelope glycoprotein of the human immunodeficiency virus type 1 (hiv-1) and a truncated variant designated gp160(t) which lacks a transmembrane domain. glycosylation, proteolytic cleavage, secretion, and biological activities of gp160 and gp160(t) 160(t) were studied in spodoptera frugiperda cells. both proteins were rapidly glycosylated and initially were found to be totally endo-beta-n-acetyl-d-glucosaminidase h (endo-h) sensitive. howe ... | 1990 | 2111957 |
| synthesis of tetanus toxin fragment c in insect cells by use of a baculovirus expression system. | the baculovirus expression vector p36c was used to express in cells of the insect spodoptera frugiperda fragment c of tetanus toxin under the control of the strong polyhedrin promoter. fragment c was expressed intracellularly at a high level and was soluble, allowing it to be purified by affinity chromatography with monoclonal antibody tt08. purified fragment c from baculovirus was used to immunize mice and was shown to successfully prevent the symptoms of tetanus following a toxin challenge. th ... | 1991 | 2019434 |
| alpha 1----3-galactosyltransferase: the use of recombinant enzyme for the synthesis of alpha-galactosylated glycoconjugates. | we have reported the isolation and characterization of a bovine cdna clone containing the complete coding sequence for udp-gal:gal beta 1----4glcnac alpha 1----3-galactosyltransferase [joziasse, d. h., shaper, j. h., van den eijnden, d. h., van tunen, a. j. & shaper, n. l. (1989) j. biol. chem. 264, 14290-14297]. insertion of this cdna clone into the genome of autographa californica nuclear polyhedrosis virus (acnpv) and subsequent infection of spodoptera frugiperda (sf9) insect cells with recom ... | 1990 | 2116309 |
| full-length cd4 electroinserted in the erythrocyte membrane as a long-lived inhibitor of infection by human immunodeficiency virus. | recombinant full-length cd4 expressed in spodoptera frugiperda 9 cells with the baculovirus system was electroinserted in erythrocyte (rbc) membranes. of the inserted cd4, 70% was "correctly" oriented as shown by fluorescence quenching experiments with fluorescein-labeled cd4. the inserted cd4 displayed the same epitopes as the naturally occurring cd4 in human t4 cells. double-labeling experiments (125i-cd4 and 51cr-rbc) showed that the half-life of cd4 electroinserted in rbc membrane in rabbits ... | 1991 | 2034680 |
| inhibition of human immunodeficiency virus type 1-induced cell fusion by recombinant human interferons. | pretreatment of hela t4 cells with recombinant alpha, beta, or gamma interferon (ifn) was found to significantly inhibit syncytium formation induced by the human immunodeficiency virus type 1 (hiv-1) envelope glycoprotein. all three ifns were found to be potent inhibitors of fusion in a system in which spodoptera frugiperda cells, infected with a baculovirus recombinant expressing the hiv-1 envelope protein, were cocultivated with hela t4 cells. in addition, these ifns were also found to block h ... | 1991 | 1920634 |
| identification of bluetongue virus vp6 protein as a nucleic acid-binding protein and the localization of vp6 in virus-infected vertebrate cells. | recently the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv) has been effectively adapted as a highly efficient vector in insect cells for the expression of various genes. a cdna sequence of rna segment 9 of bluetongue virus serotype 10 (btv-10, an orbivirus member of the reoviridae family) encoding a minor core protein (vp6) has been inserted into the bamhi site of the pacym1 transfer vector derived from acnpv. spodoptera frugiperda cells were cotransfected with the ... | 1990 | 2152806 |
| the single cysteine residue on an alpha family chick liver glutathione s-transferase cl 3-3 is not functionally important. | chick liver glutathione s-transferase cl 3-3, expressed using a baculovirus system in spodoptera frugiperda (sf9) cells, contains a single cysteine residue per subunit. this enzyme was modified with iodoacetamide. amino acid analysis indicates that 0.85 +/- 0.10 cysteine residue was modified per enzyme subunit. gst cl 3-3 modified with iodo[14c]acetamide was further digested with trypsin and the isotope-labelled fragments were isolated. the fragment containing the cysteine residue accounts for 5 ... | 1991 | 1930229 |
| sequence comparison of cellular and viral copies of host cell dna insertions found in autographa californica nuclear polyhedrosis virus. | the nucleotide sequence and structural characteristics of two nonhomologous host dna insertions of spodoptera frugiperda (fall armyworm) origin isolated from few polyhedra mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) have been determined. neither of the host insertions contain long open reading frames suggesting that cellular genes were not introduced into the viral genome. one of the host dna insertions, ifp1.6, terminated in short imperfect inverted rep ... | 1990 | 2154879 |
| purification and characterization of the recombinant extracellular domain of human nerve growth factor receptor expressed in a baculovirus system. | to obtain the large quantities of the extracellular domain of the nerve growth factor receptor (ngf-r) necessary for structural analyses, we produced this protein in the baculovirus expression system. a cdna coding for the extracellular domain of the human ngf-r was first introduced into transfer vector pvl941. recombinant baculovirus was produced by cotransfecting spodoptera frugiperda cells with the transfer vector and dna of autographa californica nuclear polyhedrosis virus. recombinant viral ... | 1990 | 2155239 |
| synthesis of bluetongue virus (btv) corelike particles by a recombinant baculovirus expressing the two major structural core proteins of btv. | the l3 and m7 genes of bluetongue virus (btv), which encode the two major core proteins of the virus (vp3 and vp7, respectively), were inserted into a baculovirus dual-expression transfer vector and a recombinant baculovirus expressing both foreign genes isolated following in vivo recombination with wild-type autographa californica nuclear polyhedrosis virus dna. spodoptera frugiperda insect cells infected with the recombinant synthesized large amounts of btv corelike particles. these particles ... | 1990 | 2157041 |
| an insertion of insect cell dna in the 81-map-unit segment of autographa californica nuclear polyhedrosis virus dna. | in this report, a transposonlike insertion of spodoptera frugiperda insect cell dna was analyzed in single-plaque isolate e of the insect baculovirus autographa californica nuclear polyhedrosis virus (acnpv). the 634-base-pair insertion is characterized by an 18-base-pair terminal inverted repeat and carries an ecori site. this additional ecori site in the 81-map-unit segment of the dna of plaque isolate e of acnpv explains the difference between the ecori restriction map of the dna from this is ... | 1990 | 2157067 |
| enzymatic activities of overexpressed herpes simplex virus dna polymerase purified from recombinant baculovirus-infected insect cells. | biochemical characterization of the herpes simplex virus (hsv) dna polymerase, a model dna polymerase and an important target for antiviral drugs, has been limited by a lack of pure enzyme in sufficient quantity. to overcome this limitation, the hsv dna polymerase gene was introduced into the baculovirus, autographa californica nuclear polyhedrosis virus, under the control of the polyhedrin promoter to give rise to a recombinant baculovirus, bp58. bp58-infected spodoptera frugiperda insect cells ... | 1990 | 2157192 |
| generation of recombinant baculovirus via liposome-mediated transfection. | baculovirus expression vectors have become a popular method of producing recombinant proteins. production of recombinant virus requires the transfection of both the native viral dna and a transfer plasmid into insect cells where recombination takes place. while several methods of transfecting insect cells exist, we have found liposome-mediated transfection to be highly efficient. here we detail the protocols and medium needed for efficient, simple transfection of spodoptera frugiperda cells. | 1991 | 1931027 |
| high-level secretion of the extracellular domain of the human growth hormone receptor using a baculovirus system. | a chemically synthesized gene (hghr-ed) coding for the extracellular domain (ed) of the human growth hormone (hgh) receptor (hghr) was inserted into the genome of autographa californica nuclear polyhedrosis virus adjacent to the polyhedrin promoter. spodoptera frugiperda cells infected with the recombinant virus secreted a protein with hgh-binding activity into the medium. the secreted 35-kda protein was purified to near homogeneity. the purified protein exhibited a high binding affinity (kd = 0 ... | 1991 | 1937047 |
| evaluation of the role of conserved his and met residues among lipoxygenases by site-directed mutagenesis of recombinant human 5-lipoxygenase. | the 5-, 12-, and 15-lipoxygenases contain a highly conserved sequence of the form his-(x)4-his-(x)4-his-(x)17-his-(x)8-his which represents a potential binding site for non heme iron to the protein. the importance of selected amino acids within this his cluster for the activity of human 5-lipoxygenase was investigated by site-directed mutagenesis using bacteria and insect cells expression systems. after single mutation of each of the 5 his residues at positions 363, 368, 373, 391, and 400 by ser ... | 1991 | 1939225 |
| biophysical studies on the morphology of baculovirus-expressed bluetongue virus tubules. | bluetongue virus tubules were purified from spodoptera frugiperda cells infected with a recombinant baculovirus containing the ns1 gene from bluetongue virus serotype 10, and expressed under control of the autographa californica nuclear polyhedrosis virus polyhedrin promoter. these tubules were subjected to a variety of chemical and physical treatments and the resulting effects on tubule morphology were examined by electron microscopy. a number of morphological similarities were noted between bl ... | 1990 | 2167940 |
| expression of the tobacco mosaic virus movement protein using a baculovirus expression vector. | a cdna clone of the tobacco mosaic virus 30k movement protein (mp) gene was constructed and introduced into an autographa californica nuclear polyhedrosis baculovirus expression vector. infection of spodoptera frugiperda cells with the vector resulted in the synthesis of low levels of mp, which was detected by anti-mp serum as two closely related species of mr approximately 34k and a third species of 32k. the authenticity of the recombinant mp was confirmed by comparison of the protein, on the b ... | 1991 | 1940871 |
| expression of avian leukaemia virus env-gp85 in spodoptera frugiperda cells by use of a baculovirus expression vector. | we studied the genetic expression of gp85 of avian leukaemia virus (alv) subgroup a in a baculovirus/insect cell system. 5'terminal sequences of the gag gene were added to precede the alv gp85 sequence and a stop codon was introduced at the boundary of gp85 and gp37. the resulting construct was then cloned into the baculovirus transfer vector pacym1, which contains the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). cells of the insect spodoptera frugiperda (sf9 ... | 1990 | 2174957 |
| expression of the full-length products of the human papillomavirus type 6b (hpv-6b) and hpv-11 l2 open reading frames by recombinant baculovirus, and antigenic comparisons with hpv-11 whole virus particles. | the l2 open reading frames (orfs) of human papillomavirus (hpv) types 6b and 11 were expressed as full-length non-fusion proteins in spodoptera frugiperda (sf-9) cells using recombinant baculovirus. both proteins were detected on western blots as immunoreactive bands which migrated with apparent mrs of 76k and 78k, respectively, and contained both cross-reactive and type-specific epitopes, as determined by polyclonal antisera directed against defined subregions of the hpv-6b and hpv-11 l2 orfs. ... | 1990 | 2174962 |
| characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system. | a cdna clone encoding the entire open reading frame of mouse entactin was constructed. this cdna was inserted into autographa californica multiple nuclear polyhedrosis virus and expressed in spodoptera frugiperda, sf9, insect cells. the recombinant entactin was produced in large quantities under the control of the polyhedrin promoter. amino terminus sequence analysis indicated that the signal peptide in the preprotein was correctly cleaved. polyclonal antibodies prepared against either the natur ... | 1990 | 2180961 |
| modulation of trehalase activity in two insect cell lines by virus infection and trehalose. | trehalase (ec 3.2.1.28), an important glycosidase involved in regulating trehalose levels and metabolic energy in insects, was measured in cell lines from fall army worm, spodoptera frugiperda and salt marsh caterpillar, estigmene acrea, treated with either glucose or trehalose in the presence or absence of tipula iridescent virus (tiv), a cytoplasmic deoxyribovirus. in medium containing 15-35 mm trehalose, both of these cells increased their trehalase activities by 4.5 to 8x the basal levels fr ... | 1991 | 1953800 |