Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| overproduction and purification of mycobacterium tuberculosis chaperonin 10. | the overexpression of the mycobacterium tuberculosis chaperonin 10 (cpn10)-encoding gene was accomplished using baculovirus expression vectors. the product was immunoreactive with a cpn10 monoclonal antibody (mab) and had an electrophoretic mobility identical to authentic cpn10. the baculovirus system was most successful in terms of reaching nearly the full expression potential of the system. recombinant cpn10 was purified from recombinant baculovirus-infected spodoptera frugiperda cells by isoe ... | 1994 | 7959041 |
| reconstitution of the recombinant 70-kda subunit of the clathrin-coated vesicle h+ atpase. | vacuolar-type proton pumps are complex heterooligomers. when dissociated into subcomplexes and subunits, the partial reactions of atp hydrolysis and transmembranous proton flow can be assigned to isolated domains. data suggest that the molecular site of atp hydrolysis resides within the 70-kda subunit but that atpase activity likely requires at least three additional subunits of 58, 40, and 33 kda (xie, x.-s., and stone, d. k. (1988) j. biol. chem. 263, 9859-9867). we have now cloned and sequenc ... | 1994 | 7961699 |
| comparative morphology of gag protein structures produced by mutants of the gag gene of human immunodeficiency virus type 1. | six mutants that differ in the extent of their carboxyterminal sequences and two deletion mutants of the gag gene of hiv-1 have been characterized morphologically following their expression in spodoptera frugiperda cells using recombinant baculoviruses. electron microscopy has revealed distinct morphological forms of the gag protein that can be classified as either (i) particulate, three-dimensional, spherical or tubular shells or (ii) non-particulate, two-dimensional, flat, curved or convoluted ... | 1994 | 7964608 |
| expression of the newcastle disease virus (ndv) fusion glycoprotein and vaccination against ndv challenge with a recombinant baculovirus. | the hybrid baculovirus constructed from autographa california nuclear polyhedrosis virus (npv) and bombyx mori (silkworm) npv was used for expression of fusion glycoprotein (f) of newcastle disease virus (ndv) strain d26. the gene encoding f protein was introduced into the improved baculovirus expression vector derived from the host-range-expanded baculovirus. in spodoptera frugiperda (fall armyworm) cells infected with a recombinant baculovirus, hyf121, the expressed f protein was properly loca ... | 1994 | 7702510 |
| suppression of apoptosis in insect cells stably transfected with baculovirus p35: dominant interference by n-terminal sequences p35(1-76). | expression of p35 from the dna genome of autographa californica nuclear polyhedrosis virus (acmnpv) suppresses virus-induced apoptosis and promotes virus replication in spodoptera frugiperda (sf21) cells. to examine the molecular mechanism by which p35 prevents apoptosis in insects, sf21 cells were stably transfected with p35. neomycin-resistant cell lines that synthesized protein p35 were identified. stable transfection with p35 protected sf21 cells from apoptosis induced by actinomycin d conce ... | 1994 | 7966562 |
| functional characterization of partially purified epstein-barr virus dna polymerase expressed in the baculovirus system. | the dna polymerase gene of epstein-barr virus (ebv) was cloned into baculovirus transfer vector (pbluebac). the recombinant baculovirus (acebp-15) was obtained by cotransfection of spodoptera frugiperda (sf9) cells with infectious dna from autographa californica multiple nuclear polyhedrin virus (acmnpv) and pbluebac plasmid carrying ebv polymerase gene. infection of sf9 cells with the recombinant virus produced substantial quantities of the ebv dna polymerase protein of the expected size (110 k ... | 1994 | 7975269 |
| expression of ricin b chain in spodoptera frugiperda. | dna encoding ricin b chain was derived from preproricin genomic dna and ligated into the baculovirus transfer vector, pacgp67a. co-transfection of spodoptera frugiperda sf9 cells with baculogold dna was followed by limiting dilution purification of recombinant baculovirus. infection of sf9 cells at a multiplicity of infection of 1 in the presence of 25 mm lactose produced 3 mg/l of soluble, glycosylated, 34 kda protein immunoreactive with monoclonal and polyclonal antibodies to ricin b chain. th ... | 1994 | 7980448 |
| overexpression, purification, and characterization of full-length and mutant caldesmons using a baculovirus expression system. | three recombinant chicken gizzard caldesmon (cad) baculovirus vectors that contained the full-length cad codon sequence (pv1cad), the full-length cad codon sequence and a six-histidine tag at the 5'-end (pbluebachiscad), or the full-length cad codon sequence and an extra six-histidine codon sequence at the 3'-end (pvlhiscad) were constructed. spodoptera frugiperda (sf9) cells transfected with these constructs overexpressed full-length cad, yielding 2, 20, and 50 micrograms per 10(6) cells for pb ... | 1994 | 7706421 |
| expression and characterization of the alpha 2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (ampa)-selective glutamate receptor channel in a baculovirus system. | using a baculovirus expression vector system, the alpha 2 subunit of the mouse alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (ampa)-selective glutamate receptor (glur) channel was expressed in spodoptera frugiperda insect cells. immunoblotting using the antibody made to the synthetic peptide corresponding to the c-terminus of glur alpha 2 and [35s]methionine/[35s]cysteine metabolic radiolabeling revealed the major 102-kda and the minor 98-kda protein bands. metabolic radiolabeling with t ... | 1994 | 7889366 |
| metallopanstimulin gene product produced in a baculovirus expression system is a nuclear phosphoprotein that binds to dna. | the protein product of the human metallopanstimulin (mps-1) gene was produced in the insect cell line spodoptera frugiperda (sf9) using the baculovirus expression vector system autographa californica nuclear polyhedrosis virus (acnpv). when a cloned mps-1 complementary dna sequence was inserted into the acnpv viral genome downstream from the promoter of the polyhedrin gene, a polypeptide with an apparent molecular weight of approximately 10,000 was observed in extracts of infected sf9 cells. thi ... | 1994 | 7986747 |
| molecular characterization of a strain-specific repeated dna sequence in the fall armyworm spodoptera frugiperda (lepidoptera: noctuidae). | the fall armyworm moth, spodoptera frugiperda, is a migratory species composed of sympatric corn and rice strains. the strains are indistinguishable in morphology but can be recognized by molecular markers. we have cloned and characterized seven monomer units of a repeated dna sequence, called fr, which is found exclusively in the genome of the rice strain individuals. the 189 bp fr units are tandemly organized in arrays longer than 30 kb. female individuals possess over 100-fold more of the fr ... | 1994 | 7987522 |
| baculovirus expression of non-structural protein ns2 and core protein vp7 of african horsesickness virus serotype 3 and their use as antigens in an indirect elisa. | non-structural protein ns2 and core protein vp7 of african horsesickness virus serotype 3 (ahsv3) were expressed in spodoptera frugiperda cells by recombinant baculoviruses containing the relevant genes. these proteins were purified and analysed by polyacrylamide gel electrophoresis and western blot. ns2 and vp7 were used separately as antigens in an indirect elisa for the detection of ahsv antibodies. both antigens cross-reacted with hyperimmune guinea-pig antisera to infected cell lysates of a ... | 1994 | 7989441 |
| iodinated 2-aminotetralins and 3-amino-1-benzopyrans: ligands for dopamine d2 and d3 receptors. | in developing selective ligands for dopamine d2 and d3 receptors, several iodinated 2-aminotetralins and 3-amino-1-benzopyrans, trans-7-hydroxy-2-[n-(3'-iodo-2'- propenyl)amino]tetralin (1), trans-monohydroxy-2-[n-propyl-n-(3'-iodo-2'-propenyl)amino]tetralin (7-, 5-, and 6-oh-pipat) (2, 3, and 4), and trans-monohydroxy-3,4-dihydro-3-[n-propyl-n-(3'-iodo-2'- propenyl)-amino]-2h-1-benzopyran (6- and 8-oh-benzopyrans) (5 and 6), were prepared. these compounds were evaluated for their binding profil ... | 1994 | 7990123 |
| cloning and production of functional active recombinant hepatitis b virus surface antigen binding protein. | endonexin ii present on the surface of human hepatocytes has recently been identified as a hepatitis b virus surface antigen (hbsag) binding protein. a full-length cdna clone encoding human endonexin ii was isolated from a human liver cdna library and was placed under the control of the polyhedrin promoter of autographa californica nuclear polyhedrosis virus (acnpv). infection of spodoptera frugiperda cells with recombinant virus resulted in the production of high amounts of recombinant protein. ... | 1994 | 7999073 |
| expression of oligohistidine-tagged ricin b chain in spodoptera frugiperda. | dna encoding adpgh6g was fused to the 5'-end of rtb dna and subcloned as a bamhi-ecori dna cassette into the baculovirus transfer vector, pacgp67a. spodoptera frugiperda sf9 cells were cotransfected with pacgp67a-adpgh6g-rtb dna and baculogold acnpv dna, and recombinant baculovirus was isolated by two cycles of limiting dilution assay followed by dot blot analysis with 32p-dctp random primer labeled rtb dna. recombinant virus was purified and amplified to obtain stocks at titers of 10(7) infecti ... | 1994 | 7533004 |
| the structure and function of the first component of complement: genetic engineering approach (a review). | the availability of cdna and genomic clones for the subcomponents of c1, as well as the recognition of the modular organization of serine-proteases have opened up exciting new possibilities for approaching structural problems. in this review the latest achievements of combined protein engineering, functional and structural studies are summarized. the concept of this research is to construct deletion, point and hybrid mutants of the highly homologous c1r and c1s subcomponents, to reveal the funct ... | 1994 | 7866721 |
| production of measles nucleoprotein in different expression systems and its use as a diagnostic reagent. | measles nucleoprotein has been successfully expressed in three different hosts, bacterial (escherichia coli bl21 de3), insect (spodoptera frugiperda; sf9) and mammalian (primary human fibroblasts) cells, each producing an antigenic protein of m(r) 60 kda. the nucleoprotein produced in all three hosts was used in an elisa for the detection of antibodies to measles virus in a cohort of haemagglutinin-positive or -negative human sera. data produced from baculovirus and adenovirus-based antigens ind ... | 1994 | 7868644 |
| enzymatic catalysis of disulfide formation. | the formation of disulfide bridges in membrane and secretory proteins occurs during the protein folding process in the endoplasmic reticulum of eukaryotic cells and the periplasm of prokaryotes. the formation of disulfide bridges is facilitated by the thiol-disulfide oxidoreductases, protein disulfide isomerase (pdi) in eukaryotes and dsba in prokaryotes. structure-function analysis of these soluble proteins demonstrates that their active sites are sequences with similarity to the active site of ... | 1994 | 7909462 |
| similar ligand binding in recombinant human alpha 2 c2-adrenoceptors produced in mammalian, insect and yeast cells. | ligand binding properties were investigated in recombinant human alpha 2c2-adrenoceptors expressed in three different host systems: shionogi s115 mouse mammary tumour cells, spodoptera frugiperda sf9 insect cells and saccharomyces cerevisiae yeast cells. the expected 43 kda alpha 2c2 protein was visualized with immunoblotting using a polyclonal alpha 2c2-receptor antibody. [3h]rauwolscine binding in cell homogenates or membranes (bmax 3-11 pmol/mg protein; kd approximately 5.5 nm) was inhibited ... | 1994 | 7911432 |
| human 17 beta-hydroxysteroid dehydrogenase: overproduction using a baculovirus expression system and characterization. | estrogenic 17 beta-hydroxysteroid dehydrogenase (17 beta-hsd) plays a pivotal role in the synthesis of estrogens. we overproduced human placental estrogenic 17 beta-hsd using a baculovirus expression system for the study of the enzyme mechanism. a cdna encoding the entire open reading frame of human 17 beta-hsd was inserted into the genome of autographa californica nuclear polyhedrosis virus and expressed in spodoptera frugiperda (sf9) insect cells. metabolic labeling and western blot analysis u ... | 1994 | 7918113 |
| synapsin iia bundles actin filaments. | synapsins are neuron-specific phosphoproteins associated with small synaptic vesicles in the presynaptic nerve terminal. synapsin i, which has been demonstrated to bundle f-actin in vitro, has been postulated to regulate neurotransmitter release by cross-linking synaptic vesicles to the actin cytoskeleton. to investigate the possible interaction of synapsin ii with actin filaments, we expressed synapsin ii in spodoptera frugiperda and high five insect cells using a recombinant baculovirus. purif ... | 1994 | 7931311 |
| expression of functional metabotropic and ionotropic glutamate receptors in baculovirus-infected insect cells. | an ionotropic glutamate receptor of the kainate subtype (glur6) and a g-protein coupled metabotropic glutamate receptor (mglur1 alpha) were expressed and studied in two insect cell lines: sf9 cells from spodoptera frugiperda and mg1 cells from trichoplusia ni. application of kainate to glur6-infected mg1 cells produced kainate-activated currents. glutamate activation of mglur1 alpha in mg1- and sf9-infected cells caused rapid, transient increases in intracellular calcium levels. this effect was ... | 1994 | 7936400 |
| cystine-rich fish antifreeze is produced as an active proprotein precursor in fall armyworm cells. | recombinant cystine-rich fish antifreeze protein (afp) was produced by fall armyworm cells from a baculovirus expression vector containing sea raven afp cdna. the natural signal sequence encoded in the cdna directed secretion of the antifreeze into the medium, from where it was recovered and purified to homogeneity. the m(r) of the exported protein (16k), as determined by sds-page, was larger than that (14k) of mature afp isolated from sea raven serum. sequencing of the recombinant afp showed th ... | 1994 | 7945337 |
| expression of a cannabinoid receptor in baculovirus-infected insect cells. | a cannabinoid receptor recombinant baculovirus (acnpv-thcr) has been constructed and employed to express rat neural cannabinoid receptors. northern analysis of total rna from spodoptera frugiperda (sf9) insect cells infected with acnpv-thcr revealed novel hyper-production of a 3.3 kb transcript when probed with nick-translated rat cannabinoid receptor cdna. optimal viral protein expression was observed in 35s-metabolically labeled acnpv-thcr-infected sf9 cells at a multiplicity of infection of 2 ... | 1994 | 7945417 |
| cloning and expression of a soluble sialidase from chinese hamster ovary cells: sequence alignment similarities to bacterial sialidases. | a cdna encoding a soluble sialidase from chinese hamster ovary (cho) cells has been cloned and expressed. completely degenerate oligonucleotide primers, which were based on the amino acid sequence of peptides obtained from the purified sialidase (warner et al., glycobiology, 3, 455-463, 1993), and the polymerase chain reaction, with single-stranded cdna template, were employed to generate a unique oligonucleotide probe. the unique probe of 93 bp was used for screening a lambda gt 10 cho cell cdn ... | 1994 | 7949662 |
| isolation and characterization of c-fos recombinant baculovirus. | the use of the baculovirus system to produce recombinant proteins is based on the high level of protein production and the possibility to obtain, in spodoptera frugiperda insect cells, recombinant proteins with the post-translational modifications found in the native proteins. here we describe the isolation and characterization of a recombinant baculovirus containing the mouse c-fos gene. the c-fos cdna was subcloned into the pvl1392 baculovirus transfer vector. the recombinant plasmid (pvl1392. ... | 1994 | 7640625 |
| functional reconstitution in lipid vesicles of influenza virus m2 protein expressed by baculovirus: evidence for proton transfer activity. | the influenza virus m2 protein was expressed from a recombinant baculovirus in spodoptera frugiperda sf9 cells, purified and reconstituted into artificial membrane vesicles. the specific inhibitor amantadine overcame the toxic activity of the protein and boosted the rate of m2 synthesis by a factor of 10, allowing yields of about 1 mg of purified m2 protein per g of sf9 cells. m2 protein expressed in this system was phosphorylated and palmitoylated and displayed properties similar to the authent ... | 1994 | 7527837 |
| characterization by electrospray mass spectrometry of human ca(2+)-sensitive cytosolic phospholipase a2 produced in baculovirus-infected insect cells. | the 85-kd cytosolic phospholipase a2 (cpla2) is a novel receptor-regulated phospholipase that is thought to initiate the production of inflammatory lipid mediators. since cpla2 is present only in minute amounts (less than 0.01% of total cellular protein) in various cells and tissues, we have used the baculovirus expression system to produce sufficient quantities of cpla2 for structural and functional analysis. the cdna for cpla2 was cloned into a baculovirus expression vector and, upon infection ... | 1994 | 7764328 |
| heterologous expression of the plant proteins strictosidine synthase and berberine bridge enzyme in insect cell culture. | the heterologous expression of cdnas encoding the alkaloid biosynthetic enzymes, strictosidine synthase [ec 4.3.3.2] from rauvolfia serpentina and the berberine bridge enzyme [(s)-reticuline: oxygen oxidoreductase (methylene-bridge-forming), ec 1.5.3.9] from eschscholtzia californica, has been achieved in a cell culture (sf9) of the fall army worm, spodoptera frugiperda, using a baculovirus-based expression system. the expression resulted in the overproduction of each plant enzyme in a catalytic ... | 1994 | 7764480 |
| optimization of the growth conditions of sf21 insect cells for high-density perfusion culture in stirred-tank bioreactors. | spodoptera frugiperda insect cells (iplb-sf21-ae) (sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. in the present study the culture conditions of these insect cells were studied to establish high-density suspension cultures with prolonged exponential growth phases. the sf21 cells were grown in 125-ml spinner flasks using five different culture media supplemented with 5% fetal c ... | 1994 | 7764890 |
| plaque assay and replication of tipula iridescent virus in spodoptera frugiperda ovarian cells. | a plaque assay was developed for the study of tipula iridescent virus (tiv) replication using a cell line derived from the fall army worm spodoptera frugiperda (sf9). infection and plaque formation were monitored with time by phase contrast microscopy, video and fluorescent light microscopy. structure of virions, viroplasmic centres and organelles of infected cells were examined by transmission electron microscopy (tem). after 4 h postinfection, plaques were visibly detected within the cell mono ... | 1994 | 7839010 |
| preparation and characterization of human rheumatoid arthritic synovial fluid phospholipase a2 produced by recombinant baculovirus-infected insect cells. | we prepared human rheumatoid arthritic synovial fluid phospholipase a2 (pla2) [ec 3.1.1.4] from insect cells infected with a recombinant baculovirus. the pla2 dna was designed, changing the original codons to those used frequently in the polyhedrin gene. sixteen oligo-deoxynucleotides ranging from 40 to 70 nucleotides were chemically synthesized and then assembled to form the whole pla2 gene. the gene thus synthesized was then placed under the control of the polyhedrin promoter of autographa cal ... | 1994 | 7798190 |
| a prototype recombinant vaccine against respiratory syncytial virus and parainfluenza virus type 3. | we have produced a genetically-engineered chimeric protein composed of the external domains of the respiratory syncytial virus (rsv) fusion (f) protein and the parainfluenza virus type 3 (piv-3) hemagglutinin-neuraminidase (hn) protein in insect cells using the baculovirus expression system. the yield of the soluble chimeric frsv-hnpiv-3 protein could be increased approximately 2-fold by using trichoplasia ni (high five) insect cells in place of spodoptera frugiperda (sf9) for expression. the ch ... | 1994 | 7765021 |
| expression, glycosylation, and phosphorylation of human keratins 8 and 18 in insect cells. | the filament forming ability and post-translational modifications of the human intermediate filaments, keratin polypeptides 8 and 18 (k8/18), were studied in recombinant baculovirus-infected insect (spodoptera frugiperda, sf9) cells. no change in cell morphology was noted after high levels of k8/18 were expressed in sf9 cells coinfected with recombinant virus-containing human k8 and k18. immunofluorescence staining showed that k8/18 expressed in sf9 cells formed somewhat what disorganized and ro ... | 1994 | 7510249 |
| expression of infectious pancreatic necrosis virus polyprotein and vp1 in insect cells and the detection of the polyprotein in purified virus. | in order to study the molecular biology of infectious pancreatic necrosis virus (ipnv) replication, six different recombinant baculoviruses were constructed. the following four recombinants contained genome segment a-specific sequences; (i) acpp contained the complete polyprotein coding region and spodoptera frugiperda (sf) cells infected by these recombinants synthesized the 106-kda polyprotein (nh2-prevp2-ns protease-vp3-cooh), which was only partially processed by the protease to yield prevp2 ... | 1994 | 7507279 |
| the alpha-5 segment of bacillus thuringiensis delta-endotoxin: in vitro activity, ion channel formation and molecular modelling. | a peptide with a sequence corresponding to the highly conserved alpha-5 segment of the cry delta-endotoxin family (amino acids 193-215 of bacillus thuringiensis cryiiia [gazit and shai (1993) biochemistry 32, 3429-3436]), was investigated with respect to its interaction with insect membranes, cytotoxicity in vitro towards spodoptera frugiperda (sf-9) cells, and its propensity to form ion channels in planar lipid membranes (plms). selectively labelled analogues of alpha-5 at either the n-terminal ... | 1994 | 7529493 |
| enzyme-linked immunosorbent assay using a recombinant baculovirus-expressed bacillus anthracis protective antigen (pa): measurement of human anti-pa antibodies. | we developed an antigen capture enzyme-linked immunosorbent assay (elisa) which does not require purified protective antigen (pa) for detection of human antibodies to bacillus anthracis pa. lysates of spodoptera frugiperda (sf-9) cells infected with recombinant baculovirus containing the pa gene were used as the source of pa to develop the elisa. recombinant pa from crude sf-9 cell lysates or pa purified from b. anthracis sterne strain was captured by an anti-pa monoclonal antibody coated onto m ... | 1994 | 7496927 |
| respiratory syncytial virus nucleocapsid protein (n) expressed in insect cells forms nucleocapsid-like structures. | the gene coding for the n protein of rsv strain long has been cloned and sequenced. it was introduced behind the polyhedrin promoter of the shuttle vector pvl941 and baculoviruses containing the n gene were constructed by homologous recombination. infection of spodoptera frugiperda 9 cells resulted in the production of large amounts of a protein similar in size and antigenicity to the authentic n protein. the baculovirus expressed n protein was concentrated in the cytoplasm of the insect cells a ... | 1994 | 7513931 |
| desensitization, phosphorylation and palmitoylation of the human dopamine d1 receptor. | the regulation and post-translational modifications of the human dopamine d1 receptor were studied in the baculovirus-eukaryotic cell expression system. baculovirus constructs containing either the dna encoding the dopamine d1 receptor or a dna encoding a c-myc epitope tagged dopamine d1 receptor (c-myc-dopamine d1 receptor) were used to infect spodoptera frugiperda (sf9) insect cells. expressed dopamine d1 and c-myc-dopamine d1 receptors bound agonists and antagonists with affinities and a rank ... | 1994 | 7515822 |
| unprocessed foot-and-mouth disease virus capsid precursor displays discontinuous epitopes involved in viral neutralization. | a foot-and-mouth disease virus (fmdv) cdna cassette containing sequences encoding the capsid precursor p1, peptide 2a and a truncated 2b (abbreviated p1-2a) of type c fmdv, has been modified to generate the authentic amino terminus and the myristoylation signal. this construct has been used to produce a recombinant baculovirus (acmm53) which, upon infection of spodoptera frugiperda insect cells, expressed a recombinant p1-2a precursor with a high yield. this polyprotein reacted with neutralizing ... | 1994 | 7515974 |
| high-level expression of functional glutamate receptor channels in insect cells. | we have expressed glutamate-gated ion channels in spodoptera frugiperda sf21 insect cells using a recombinant baculovirus system. cells infected with recombinant baculoviruses encoding the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (ampa)-selective glutamate receptor channel subunits glur-b and glur-d displayed specific high-affinity [3h]ampa binding (apparent dissociation constant kd of 15 nm for glur-b and 40 nm for glur-d) with pharmacological profiles typical of ampa receptors. the ... | 1994 | 7519022 |
| ligand-binding properties and n-glycosylation of alpha 1 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate(ampa)-selective glutamate receptor channel expressed in a baculovirus system. | the alpha 1 subunit of the mouse alpha-amino-3-hydroxy-5-methyl-4-isoxazole- propionate(ampa)-selective glutamate receptor channel has been expressed in insect spodoptera frugiperda cells using a baculovirus system. the recombinant receptor proteins were identified by immunocytochemical detection, western-blot analysis, and [35s]methionine/[35s]cysteine metabolic labeling experiments. the effect of tunicamycin on the metabolic labeling and immunoblots suggested that the two products, a major pro ... | 1994 | 7519985 |
| characterization of the spodoptera frugiperda tata-binding protein: nucleotide sequence and response to baculovirus infection. | a cdna clone containing a 921 bp open-reading frame (307 amino acids; 34 kda) homologous to the tata-binding protein (tbp) was isolated and sequenced from a spodoptera frugiperda cell line that is commonly used in the baculovirus expression system. analysis of the s. frugiperda tbp (sftbp) sequence showed that the amino-terminal portion of sftbp diverged significantly from that of other tbp sequences including drosophila melanogaster whereas the carboxy-terminal sequence was highly conserved. so ... | 1994 | 7520800 |
| fkbp46, a novel sf9 insect cell nuclear immunophilin that forms a protein-kinase complex. | recently, we identified a 59-kda nuclear phosphoprotein that is associated with a recombinant mouse fkbp-52 (alnemri, e. s., fernandes-alnemri, t., nelki, d. s., dudley, k., dubois, g. c., and litwack, g. (1993) proc. natl. acad. sci. u.s.a. 90, 6839-6843). here we describe the cloning, overexpression, and characterization of this protein from spodoptera frugiperda insect cells (sf9 cells). the cloned cdna codes for an acidic protein of 412 amino acids with distinct structural domains. starting ... | 1994 | 7527037 |
| characterization of neuronal nitric oxide synthase and a c415h mutant, purified from a baculovirus overexpression system. | nitric oxide synthase (nos) catalyzes the conversion of l-arginine to citrulline and nitric oxide (.no). a baculovirus overexpression system has been developed for a constitutive nos isoform, cloned originally from rat cerebellum (b-nos). recombinant virus was used at a multiplicity of infection of 5 to infect spodoptera frugiperda cells in culture, and nos was expressed to 10% of the total soluble protein at 48 h postinfection. in order to express catalytically active enzyme, it was necessary t ... | 1994 | 7527656 |
| reconstitution of the influenza virus m2 ion channel in lipid bilayers. | m2, an integral membrane protein of influenza a virus, was purified from either influenza a virus-infected cv-1 cells or from spodoptera frugiperda (sf9) cells infected with a recombinant-m2 baculovirus. the purified protein, when incorporated into phospholipid bilayer membranes, produced ion-permeable channels with the following characteristics: (1) the channels appeared in bursts during which unit conductances of diverse magnitudes (25-500 ps) were observed. (2) the most probable open state wa ... | 1994 | 7535851 |
| quantification of damage to suspended insect cells as a result of bubble rupture. | it is proposed that when cells are either attached to, or very near, a rupturing bubble, the hydrodynamic forces associated with the rupture are sufficient to kill the cells. four types of experiments were conducted to quantify the number and location of these killed cells. we determined: (1) the number of cells killed as a result of a single, 3.5-mm bubble rupture; (2) the number and viability of cells in the upward jet that results when a bubble ruptures; (3) the number of cells on the bubble ... | 1994 | 18613308 |
| evaluation of the spatial pattern of steinernema riobravis in corn plots. | the vertical and horizontal spatial patterns of a naturally occurring population of the entomopathogenic nematode steinernema riobravis (rhabditida: steinernematidae) were investigated in corn field soil by laboratory and field bioassays. this nematode appears to be endemic to the lower rio grande valley of texas, where it was found parasitizing prepupae and pupae of both corn earworm, helicoverpa zea, and fall armyworm, spodoptera frugiperda (lepidoptera: noctuidae). corn earworm prepupa was th ... | 1994 | 19279865 |
| baculovirus expression system scaleup by perfusion of high-density sf-9 cell cultures. | a perfusion system based on a 4-l stirred tank bioreactor and a custom-designed tangential (cross-flow) filter was assembled to realize a scaleup of the baculovirus expression vector system (bevs). when perfused with 1 to 1.5 vol/day, spodoptera frugiperda (sf-9) insect cell cultures grew from 4 x 10(6) to 15 x 10(6) cells/ml over 3 to 4 days. the possibility of maintaining high specific production of recombinant vp6 protein (from bovine rotavirus) after baculovirus infection of the high-density ... | 1994 | 18615881 |
| the effect of oscillating dissolved oxygen concentrations on the metabolism of a spodoptera frugiperda iplb-sf21-ae clonal isolate. | the effect of oscillating dissolved oxygen (do) concentration on the metabolism of a clonal isolate of the spodoptera frugiperda iplb-sf21-ae insect cell line was investigated. specifically, the effect on cell growth, re- combinant protein synthesis, glucose and glutamine consumption, and lactate accumulation was determined. prior to conducting the oscillating do experiments, it was found that the do concentration could be reduced to 15% air saturation without adversely affecting the growth rate ... | 1995 | 18623445 |
| pp60src-mediated phosphorylation of connexin 43, a gap junction protein. | several laboratories have demonstrated a decrease in gap junctional communication in cells transformed by the src oncogene of the rous sarcoma virus. the decrease in gap junctional communication was associated with tyrosine phosphorylation of the gap junction protein, connexin 43 (cx43). this study was initiated to determine if the phosphorylation of cx43 is the result of a direct kinase-substrate interaction between the highly active tyrosine kinase, pp60v-src, and cx43. previous biochemical st ... | 1995 | 7539006 |
| the impact of three-dimensional structure on the expression of pla alloantigens on human integrin beta 3. | we have compared the binding of affinity-purified anti-pla1 igg from seven nonrelated donors with chimeric integrin subunit beta 3 molecules expressed in the baculovirus-spodoptera frugiperda insect cell system. beta 3 chimeras were engineered to include segments of antigenic human beta 3 sequences spliced to intervening segments of nonantigenic xenopus beta 3 sequence. our results clearly show that antibodies from all seven donors will bind to nondenatured molecules containing the antigenic hum ... | 1995 | 7540884 |
| the exchange of arg-gly-asp (rgd) and arg-tyr-asp (ryd) binding sequences in a recombinant murine fab fragment specific for the integrin alpha iib beta 3 does not alter integrin recognition. | the murine monoclonal antibody opg2 is an excellent paradigm of natural rgd ligands and binds specifically to alpha iib beta 3 integrin. a reactive arg103-tyr104-asp105 (ryd) tripeptide is located in an extended loop, the third complementarity-determining region of the heavy chain (h3). when compared to other rgd ligands, the ryd tripeptide of opg2 is unique, in that the side chains are fixed in a stable orientation that we have defined by x-ray crystallography. in this study, we express opg2 h ... | 1995 | 7542651 |
| two domains with amino-acid sequence similarity are required for dihydroneopterin aldolase function in the multifunctional folic acid synthesis fas protein of pneumocystis carinii. | the folic acid synthesized gene (fas) of pneumocystis carinii (pc) codes for a multifunctional enzyme (fas) known to catalyse three consecutive steps leading to the production of dihydropteroate in the de novo folate synthesis pathway. previously, we predicted that a domain, designated fasb (amino acids (aa) 161-280), of the 740-aa multifunctional protein contains the first of the three enzyme activities in the pathway, namely dihydroneopterin aldolase (dhna), since it shares 23% aa identity wit ... | 1995 | 7543066 |
| cloning and expression of a cdna encoding bovine retinal pigment epithelial 11-cis retinol dehydrogenase. | identification of a 32-kd protein in the bovine retinal pigment epithelium. | 1995 | 7544779 |
| stable expression of maize auxin-binding protein in insect cell lines. | to achieve continuous expression of the major maize auxin-binding protein (abp1) in insect cells, the abp1 gene coding region was placed under control of a baculovirus immediate-early gene promoter and transfected into spodoptera frugiperda sf9 cells. the abp1 gene was detected in twelve cell lines, one of which was selected for detailed analysis. immunolocalisation demonstrated that abp1 was targeted to and retained in the endoplasmic reticulum (er), in accordance with its signal peptide and ca ... | 1995 | 7556613 |
| functional expression and segmental localization of rat colonic k-adenosine triphosphatase. | a putative cdna for the colonic k-atpase has recently been cloned (crowson, m.s., and g. e. shull. 1992. j. biol. chem. 267:13740-13748). considerable evidence exists that there are two k-atpases and active k absorptive processes in the rat distal colon: one that is ouabain sensitive and the other ouabain insensitive. the present study used the baculovirus expression system to express k-atpase activity in insect spodoptera frugiperda (sf 9) cells and a polyclonal antibody (m-1), developed agains ... | 1995 | 7560093 |
| biochemical analysis of the n-glycosylation pathway in baculovirus-infected lepidopteran insect cells. | the baculovirus-insect cell system is used routinely for foreign glycoprotein production, but the precise nature of the n-glycosylation pathway in this system remains unclear. some studies indicate that these cells cannot process n-linked oligosaccharides to complex forms containing outer-chain galactose and sialic acid, while others indicate that they can. in this study, we used the major virion envelope glycoprotein of the baculovirus autographa california multicapsid nuclear polyhedrosis viru ... | 1995 | 7571420 |
| replication of orgyia pseudotsugata baculovirus dna: lef-2 and ie-1 are essential and ie-2, p34, and op-iap are stimulatory genes. | a transient dna replication assay was used to identify genes located within m.u. 90.5-7.0 of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome that influenced replication of a reporter plasmid containing an opmnpv origin of replication, when cotransfected into uninfected lymantria dispar cells. the viral transactivator ie-1 and a 2.4-kb subclone were found to be essential for replication. the 2.4-kb region was sequenced and open reading frames were identified. ... | 1995 | 7571435 |
| evidence for proteolytic processing of tobacco mosaic virus movement protein in arabidopsis thaliana. | two ecotypes of arabidopsis thaliana were transformed with the gene encoding tobacco mosaic virus (tmv) movement protein (p30). p30 accumulated largely in a subcellular fraction containing cell wall components and as a soluble protein. the protein migrated in denaturing gels with an m(r) of 30k, significantly faster than p30 (m(r) approximately 34k) accumulating after expression in transgenic tobacco, escherichia coli or spodoptera frugiperda cells, or after virus multiplication in tobacco. the ... | 1995 | 7579611 |
| cdna-directed expression of human cytochrome p450 cyp1a1 using baculovirus. purification, dependency on nadph-p450 oxidoreductase, and reconstitution of catalytic properties without purification. | a recombinant baculovirus containing the human cytochrome p450 (cyp) 1a1 cdna was constructed and used to express cyp1a1 in spodoptera frugiperda (sf9) insect cells (0.14 +/- 0.04 nmol/mg protein, 53 +/- 14 nmol/liter, n = 30). the enzyme represented approximately 1% of total cellular protein and was partially purified by a three-column procedure to a specific content of 5.0 nmol/mg protein. catalytic activity was reconstituted with both the purified enzyme using lipid and nadph-p450 oxidoreduct ... | 1995 | 7587956 |
| [direct expression of the pig testicular 3 alpha/beta(20 beta)- hydroxysteroid dehydrogenase cdna using baculovirus expression system]. | the direct expression of the pig testicular 3 alpha/beta(20 beta)- hydroxysteroid dehydrogenase (hsd) cdna using a baculovirus expression system was investigated. a minimum essential cdna containing the coding region of 3 alpha/beta(20 beta)-hsd was amplified by polymerase chain reaction (pcr) to extend the dna linker including the bam hi restriction site in the both ends of the cdna. as the template, 3 alpha/beta(20 beta)-hsd cdna, pbs 52, which was subcloned to bluescript ii was used. the pcr ... | 1995 | 7602499 |
| expression and characterization of the zeta 1 subunit of the n-methyl-d-aspartate (nmda) receptor channel in a baculovirus system. | using a baculovirus expression vector system, the zeta 1 subunit of the mouse n-methyl-d-aspartate (nmda) receptor channel was expressed in spodoptera frugiperda insect cells. the peptide corresponding to the c-terminus of the zeta 1 subunit was synthesized by using the multiple antigen peptide (map) system, and an antibody to the synthetic peptide was produced. immunoblotting using the newly developed antibody revealed the major 122-kda and the minor 104-kda protein bands. the effect of tunicam ... | 1995 | 7609635 |
| production and purification of recombinant african swine fever virus attachment protein p12. | the conditions for cultivation of spodoptera frugiperda (sf9) insect cells for production of recombinant baculoviruses have been studied, to scale-up and improve the efficiency of the process for production of the african swine fever virus attachment protein p12 in the baculovirus expression system. it was shown that the total virus and recombinant protein production in insect cells infected with the acp12 recombinant baculovirus were slightly dependent on cell density, but largely dependent on ... | 1995 | 7612242 |
| molecular structure and function of cd4 on murine egg plasma membrane. | in the present study, the expression of the cd4 molecule on murine egg plasma membrane was confirmed by the indirect immunofluorescence (iif) method. the full-length cd4 cdna from murine eggs was synthesised by the reverse transcriptase-polymerase chain reaction (rt-pcr) method and its authenticity verified by southern blot hybridisation using an end-labelled internal oligonucleotide. the results of dna sequencing showed that the nucleotide sequence of the cdna of cd4 from murine egg mrna was id ... | 1995 | 7613876 |
| ligand binding properties of muscarinic acetylcholine receptor subtypes (m1-m5) expressed in baculovirus-infected insect cells. | five subtypes of muscarinic acetylcholine receptors (m1-m5) have been expressed in insect cells (spodoptera frugiperda, sf9) using the baculovirus system. up to 6 nmol of muscarinic acetylcholine receptors were produced by 1 liter culture; 0.3 to 0.6 (human m1), 3 to 6 (human m2), 2 to 4 (rat m3), 1 to 2 (rat m4) and 0.5 to 1 (human m5) nmol. pirenzepine, af-dx116 and hexahidrosiladifenidol showed the highest affinity for the m1, m2 and m3 subtype, respectively, indicating that these receptors e ... | 1995 | 7616422 |
| nucleotide labeling and reconstitution of the recombinant 58-kda subunit of the vacuolar proton-translocating atpase. | evidence suggests that atp hydrolysis catalyzed by the clathrin-coated vesicle proton-translocating atpase requires at least four polypeptides of molecular masses of 70, 58, 40, and 33 kda (xie, x.-s., and stone, d.k. (1988) j. biol. chem. 263, 9859-9867). to further investigate the subunit requirements for atp hydrolysis, histidine-tagged, 58-kda polypeptide was expressed in insect sf9 (spodoptera frugiperda) cells. after purification by ni(2+)-nitrolotriacetic acid chromatography, the 58-kda p ... | 1995 | 7622510 |
| agonist-induced desensitization of dopamine d1 receptor-stimulated adenylyl cyclase activity is temporally and biochemically separated from d1 receptor internalization. | the regulation of the dopamine d1 receptor was investigated by using c-myc epitope-tagged d1 receptors expressed in sf9 (fall armyworm ovary) cells. treatment of d1 receptors with 10 microm dopamine for 15 min led to a loss of the dopamine-detected high-affinity state of the receptor accompanying a 40% reduction in the ability of the receptor to mediate maximal dopamine stimulation of adenylyl cyclase activity. after 60 min of agonist exposure, 45 min after the occurrence of desensitization, 28% ... | 1995 | 7479745 |
| immune protection conferred by the baculovirus-related glycoprotein of thogoto virus (orthomyxoviridae). | the coding region of segment 4 of thogoto (tho) virus, a tick-borne member of the orthomyxoviridae, was expressed in a baculovirus system under the control of the polyhedrin promoter. this construct expressed authentic envelope glycoprotein as determined by size and antigenic reactivity with a panel of monoclonal antibodies (mabs). immunization of hamsters with spodoptera frugiperda (sf21) cells infected with the recombinant baculovirus induced neutralizing and protective antibodies against viru ... | 1995 | 7483270 |
| high level production and localization of bovine rod cgmp-gated cation channel subunit 1 in baculovirus-infected insect cells and saccharomyces cerevisiae. | the cgmp-gated cation channel is responsible for the last step in the vertebrate phototransduction cascade which couples light activation of rhodopsin to a change in membrane permeability. two different expression systems, baculovirus-infected spodoptera frugiperda (sf9) insect cells and saccharomyces cerevisiae, were used for the overproduction of the cgmp-gated cation channel subunit 1 of bovine rod cells. presence of recombinant channel protein was monitored by sds-page and western-blot analy ... | 1995 | 7488067 |
| modulation of cd4 lateral mobility in intact cells by an intracellularly applied antibody. | this study shows that the lateral mobility of cd4, an important plasma-membrane immune receptor, can be modulated by intracellular application of an anti-cd4 antibody. for this purpose, (i) full-length cd4 and a truncated cd4 mutant, lacking a 32-residue-long c-terminal intracellularly exposed domain, were expressed in spodoptera frugiperda (sf9) insect cells, (ii) a monoclonal antibody, c6, with specificity for the c-terminal domain was generated, and (iii) a versatile apparatus for fluorescenc ... | 1995 | 7492321 |
| delta-endotoxins induce cation channels in spodoptera frugiperda brush border membranes in suspension and in planar lipid bilayers. | membrane potential measurements using a fluorescent dye indicated that two specific toxins active against spodoptera frugiperda larvae (cryic and cryid) cause immediate permeability changes in midgut epithelial brush border membrane vesicles (bbmv). the initial response and the sustained permeability change are cationic, not very k+ selective, and occur at in vivo lethal doses (nm). the toxin response has a different ion selectivity and is more sensitive to ba2+ than the intrinsic cation permeab ... | 1995 | 7533728 |
| n-linked glycosylation of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (ampa)-selective glutamate receptor channel alpha 2 subunit is essential for the acquisition of ligand-binding activity. | the n-linked glycosylation of the alpha 2 subunit of the mouse alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (ampa)-selective glutamate receptor (glur) channel was characterized. the receptor subunit protein has five putative n-glycosylation sites. the recombinant receptor proteins were identified by [35s]methionine/[35s]cysteine metabolic labeling, western blot analysis, immunocytochemical detection, and [3h]ampa binding experiments when expressed in insect spodoptera frugiperda cells us ... | 1995 | 7532209 |
| construction and adhesive properties of a soluble madcam-1-fc chimera expressed in a baculovirus system: phylogenetic conservation of receptor-ligand interaction. | madcam-1 is a high endothelial venule adhesion molecule composed of immunoglobulin and mucin-like domains which binds the leucocyte integrin lpam-1 (alpha 4 beta 7), and is largely responsible for the selective homing of lymphocytes to mucosal tissues. a novel soluble form of mouse madcam-1 which is normally membrane bound has been produced by joining the extracellular region of the receptor to the fc domain of human igg1. the madcam-1-fc cdna was inserted into the genome of autographa californi ... | 1995 | 7631157 |
| comparison of the enzymatic properties of the na,k-atpase alpha 3 beta 1 and alpha 3 beta 2 isozymes. | the coexpression of multiple isoforms of the alpha and beta subunits of the na,k-atpase in mammalian tissues gives rise to the complex molecular heterogeneity that characterizes the na pump. the expression of the different na,k-atpase isoforms in insect cells using recombinant baculoviruses represents a useful system for the analysis of na,k-atpase isoform function. in the present study, we use this system to direct the expression of the rat na,k-atpase alpha 3 beta 1 and alpha 3 beta 2 in sf-9 ... | 1995 | 7632689 |
| baculovirus-driven expression and purification of glycine receptor alpha 1 homo-oligomers. | the glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. we expressed a homo-oligomeric receptor composed of human alpha 1 subunits in spodoptera frugiperda cells by infection with a recombinant autographa californica nuclear polyhedrosis virus. a substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4- to 30-fold higher than in other eukaryotic heterologous expression ... | 1995 | 7635206 |
| expression of pe38 and ie2, viral members of the c3hc4 finger family, during baculovirus infection: pe38 and ie2 localize to distinct nuclear regions. | the pe38 gene of autographa californica nuclear polyhedrosis virus represents one of the major early transcripts after viral infection. the function of the pe38 protein, which contains a c3hc4 zinc finger motif, is still not understood. we have raised polyclonal antiserum against the pe38 protein, pe38, produced in bacteria to investigate pe38 expression in the course of infection. a approximately 38-kda polypeptide is first detectable at 2 h postinfection and decreases rapidly after 24 h. durin ... | 1995 | 7636971 |
| the rotavirus nonstructural glycoprotein nsp4 mobilizes ca2+ from the endoplasmic reticulum. | we previously reported that expression of rotavirus nonstructural glycoprotein nsp4 is responsible for an increase in cytosolic free ca2+ concentration ([ca2+]i) in spodoptera frugiperda (sf9) insect cells (p. tian, y. hu, w. p. schilling, d. a. lindsay, j. eiden, and m. k. estes, j. virol. 68:251-257, 1994). the purpose of the present study was to determine the mechanism by which nsp4 causes an increase in [ca2+]i by measuring the permeability of the cytoplasmic and endoplasmic reticulum (er) m ... | 1995 | 7637021 |
| differential transcription of baculovirus late and very late promoters: fractionation of nuclear extracts by phosphocellulose chromatography. | an in vitro transcription system based on cytidine-free cassette was developed for the late 39k gene and the very late polyhedrin gene of autographa californica nuclear polyhedrosis virus (acnpv). optimization of transcription conditions revealed that a preincubation step was not required for transcription of late and very late promoters, although preincubation was essential for efficient transcription from an early promoter. the 39k and polyhedrin constructs were actively transcribed by nuclear ... | 1995 | 7707516 |
| horseradish peroxidase phe172-->tyr mutant. sequential formation of compound i with a porphyrin radical cation and a protein radical. | a gene coding for the f172y mutant of horseradish peroxidase isozyme c (hrp) has been constructed and expressed in both spodoptera frugiperda (sf-9) and trichoplusia ni egg cell homogenate (highfive) cells. homology modeling with respect to three peroxidases for which crystal structures are available places phe172 on the proximal side of the heme in the vicinity of porphyrin pyrrole ring c. the ph optimum and spectroscopic properties of the f172y mutant are essentially identical to those of wild ... | 1995 | 7629167 |
| comparison of the non-structural protein, ns1, of tick-borne and insect-borne orbiviruses. | the nucleotide sequence of rna segment 6 of broadhaven virus (brdv), a tick-borne orbivirus, was determined principally from two overlapping cdna clones and rna end sequence analysis. the genome segment is 1714 base pairs in length and has a coding capacity for a protein of 537 amino acids, having a net charge of +4.0 at neutral ph. comparison of the predicted amino acid sequence of brdv rna segment 6 with the ns1 sequence of insect-borne orbiviruses, bluetongue virus (btv), african horse sickne ... | 1995 | 7653106 |
| differential requirements for baculovirus late expression factor genes in two cell lines. | a plasmid library of 18 late expression factor (lef) genes (lef library) from the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) supports transient expression from a late viral promoter in the sf-21 cell line, derived from spodoptera frugiperda. we found, however, that this lef library was unable to support expression from the same promoter in the tn-368 cell line, derived from trichoplusia ni, which is also permissive for acmnpv replication. to identify the additional fa ... | 1995 | 7666527 |
| high-level secretion of native rat pancreatic lithostathine in a baculovirus expression system. | we have constructed a recombinant baculovirus expression vector containing rat pancreatic lithostatin cdna. baculovirus infection of spodoptera frugiperda (sf9) insect cells resulted in the de novo synthesis and secretion of a recombinant protein demonstrating an apparent molecular weight of about 16.5 kda. under optimal conditions [multiplicity of infection of 5 plaque-forming units (pfu)/cell and culture times of 48-56 h postinfection] recombinant protein was secreted into the culture medium a ... | 1995 | 7667245 |
| morphological changes of the insect cells in the baculovirus system as a function of v-myb and c-myb inserts expression and topographic localization of v-myb and c-myb proteins. | structural changes of insect cells spodoptera frugiperda in the baculovirus expression system after expression of v-myb oncogene and c-myb protooncogene inserts were studied by means of electron microscopy. expression of v-myb gene insert was accompanied by extensive changes in cell structure, when compared with those of the noninfected and wild-type virus-infected cells. enormous increase in nuclear content was apparent within 48 hrs after infection, along with changes in nucleolus appearance. ... | 1995 | 7676943 |
| nucleotide sequence and transcriptional analysis of the gp41 gene of spodoptera frugiperda nuclear polyhedrosis virus. | the spodoptera frugiperda multiple nucleocapsid nuclear polyhedrosis virus (sfmnpv) gp41 structural protein gene was located in the 1.9 kbp ecori-s fragment and sequenced. an open reading frame (orf) of 999 nucleotides was detected that encoded a protein of 332 amino acids. the gp41 gene transcript was detected after 12 h post-infection (p.i.) and remained detectable at 48 h p.i. two major mrnas, about 1.6 and 2.8 kb in length, were determined by northern blot analysis. primer extension analysis ... | 1995 | 7782772 |
| cyp3a4 expressed by insect cells infected with a recombinant baculovirus containing both cyp3a4 and human nadph-cytochrome p450 reductase is catalytically similar to human liver microsomal cyp3a4. | human cytochrome cyp3a4 is the most abundant of all the p450s in human liver and is involved in the metabolism of many environmental toxicants and drugs. kinetic studies with cyp3a4 have been hampered due to low activity of this enzyme obtained from recombinant gene expression systems or difficulty in reconstituting activity with the native enzyme purified from human liver. to overcome these obstacles, we have expressed high levels of catalytically active cyp3a4 and human nadph-cytochrome p450 r ... | 1995 | 7771780 |
| n-glycosylated polypeptides synthesized in the early phase of acnpv infected insect cells. | infected cell specific polypeptides (icsp) in autographa californica nuclear polyhedrosis virus (acnpv) infected spodoptera frugiperda cell line, sf9, were radiolabelled with [14c] mannose and the labelled proteins were monitored at various time intervals. a major glycoprotein of molecular size of 64 kda appears as early as 3h post infection (p.i.) which peaks at 12h p.i., but in presence of tunicamycin this protein is not labelled. two other glycoproteins of molecular size 67 kda and 63 kda app ... | 1995 | 7773189 |
| kinetic properties of the alpha 2 beta 1 and alpha 2 beta 2 isozymes of the na,k-atpase. | the presence of multiple isoforms of the alpha and beta subunits of the na,k-atpase in most mammalian tissues has hindered the understanding of the roles of the individual isoforms in directing na,k-atpase function. expression of the na,k-atpase subunits in insect cells using recombinant baculoviruses has proven to be a useful system for the study of na,k-atpase function. using this system, we have expressed the rat na,k-atpase alpha 2 beta 1 and alpha 2 beta 2 isoforms in sf-9 insect cells, a c ... | 1995 | 7819213 |
| characterization of an insect cell-derived theileria parva sporozoite vaccine antigen and immunogenicity in cattle. | previous data showed that six out of a group of nine cattle inoculated with ns1-p67, a recombinant form of a 67-kda theileria parva sporozoite surface protein, were immune to east coast fever. this bacterially expressed antigen encoded all 709 amino acid residues of p67 fused to the c-terminal end of 87 residues derived from ns1, a structural protein of influenza virus, and a linker dna sequence. ns1-p67 lacked reactivity with tpm 12, a monoclonal antibody to native p67, and had an estimated mol ... | 1995 | 7822014 |
| potassium channel structure and function as reported by a single glycosylation sequon. | inwardly rectifying k+ channels (irks) are highly k(+)-selective, integral membrane proteins that help maintain resting the membrane potential and cell volume. integral membrane proteins as a class are frequently n-glycosylated with the attached carbohydrate being extracellular and perhaps modulating function. however, dynamic effects of glycosylation have yet to be demonstrated at the molecular level. romk1, a member of the irk family is particularly suited to the study of glycosylation because ... | 1995 | 7797521 |
| functional chicken gizzard heavy meromyosin expression in and purification from baculovirus-infected insect cells. | the heavy chain and the essential and the regulatory light chains of chicken gizzard heavy meromyosin (hmm) were coexpressed in spodoptera frugiperda (fall armyworm) cells infected with a mixture of two recombinant autographa californica baculoviruses. soluble hmm consisting of the heavy chain and the two types of light chains was obtained. the recombinant hmm was purified from the virus-infected cells and characterized. the regulatory light chain of the isolated recombinant hmm was phosphorylat ... | 1995 | 7846039 |
| signal transduction in sf9 insect cells: endocytosis of recombinant cd4 after phorbol ester treatment. | in spodoptera frugiperda (sf9) insect cells infected with a recombinant baculovirus carrying a gene construct encoding human cd4 endocytosis of cd4 is induced after stimulation with phorbol-12-myristate-13-acetate (pma). stimulation of endocytosis with pma reduced the amount of full-length cd4 on the plasma membrane of sf9 cells by 50% after 2 hours. endocytosis of cd4 is blocked after intracellular delivery by cationic liposomes of a monoclonal antibody directed against a cytoplasmic sequence o ... | 1995 | 7857297 |
| transcriptional regulation of cell line-dependent, baculovirus-mediated expression of foreign genes. | cell line-dependent expression of foreign genes in the baculovirus system was investigated using a recombinant vac beta hcg-luc virus carrying two reporter genes--beta subunit of human chorionic gonadotropin (beta hcg) and luciferase (luc)--placed under the transcriptional control of the autographa californica nuclear polyhedrosis virus (acnpv) polyhedrin gene promoters. five different lepidopteran cell lines derived from spodoptera frugiperda (sf21 and sf9), bombyx mori (bmn and bm5), and trich ... | 1995 | 7530452 |
| molecular cloning and expression of catrocollastatin, a snake-venom protein from crotalus atrox (western diamondback rattlesnake) which inhibits platelet adhesion to collagen. | a 50 kda protein that inhibits platelet adhesion to collagen has been isolated from snake venom of crotalus atrox (western diamondback rattlesnake) and has been named 'catrocollastatin'. the cdna cloning of catrocollastatin has been accomplished. a full-length cdna of 2310 bp with an open reading frame between nucleotides 51 and 1880 was obtained. the deduced amino acid sequence consists of 609 amino acids. the cdna-predicted amino acid sequence is highly similar to that of haemorrhagic metallop ... | 1995 | 7733877 |
| functional expression of a diuretic hormone receptor in baculovirus-infected insect cells: evidence suggesting that the n-terminal region of diuretic hormone is associated with receptor activation. | a recombinant baculovirus containing the diuretic hormone receptor cdna from manduca sexta was constructed. when spodoptera frugiperda (sf9) cells were infected with the virus, a time-dependent expression of the receptor appeared. the expressed receptor displayed high affinity for diuretic hormone, which was similar to the affinity observed in malpighian tubules and transfected cos-7 cells. the receptor expression level was 77 pmol/mg protein, as compared to 3.1 pmol/mg protein in malpighian tub ... | 1995 | 7742837 |
| the autographa californica nuclear polyhedrosis virus homologous region 1a: identical sequences are essential for dna replication activity and transcriptional enhancer function. | hr1a consists of two 30-bp imperfect palindrome sequences separated by 58 bp and each palindrome contains a naturally occurring ecori site at its core. plasmid subclones of the hr1a-containing acmnpv hindiii-n fragment were examined for their ability to replicate in virus-infected (spodoptera frugiperda) sf9 cells, and to stimulate transcription when linked in cis with a 39k gene promoter-beta-glucuronidase fusion and cotransfected into cells along with a plasmid (ple-1) containing the gene enco ... | 1995 | 7747446 |
| cloning and expression of the nucleoprotein of peste des petits ruminants virus in baculovirus for use in serological diagnosis. | peste des petits ruminants (ppr) is a viral disease of goats and sheep characterized by erosive stomatitis, enteritis, and pneumonia. the virus is a member of the family paramyxoviridae and the genus morbillivirus. the disease has high morbidity and mortality rates and has a substantial economic impact in developing countries. we have cloned and sequenced the cdna of the nucleocapsid (n) gene of the nigeria 75/1 strain of ppr virus (pprv). a comparison of its nucleotide and deduced amino acid se ... | 1995 | 7747450 |
| synthesis and processing of the rubella virus p110 polyprotein precursor in baculovirus-infected spodoptera frugiperda cells. | in order to study the processing of rubella virus (rv) structural proteins (capsid protein, of 33 kda; e2 of 42-47 kda; and e1 of 58 kda) in spodoptera frugiperda (fall armyworm) cells, a 24s cdna encoding the polyprotein precursor, p110, was inserted under the transcriptional regulation of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed during viral infection. by immunoblot analysis using antibodies directed against whole rv and the in ... | 1995 | 7754676 |
| expression of trypanosoma congolense antigens in spodoptera frugiperda insect cells. | transcripts which encode two metacyclic-form-specific variable surface glycoproteins (mvsgs) of trypanosoma congolense il3000 have been cloned into baculovirus expression vectors using a novel transfer vector, pacl11. one of the recombinant baculoviruses (acvsg1) expressed a mvsg as a glycoprotein with a signal peptide which was cleaved in this expression system, whereas the other one (acvsg2) expressed an unprocessed protein. from 1 liter of culture containing 10(9) spodoptera frugiperda cells ... | 1995 | 7758544 |
| protection of guinea-pigs from experimental rocky mountain spotted fever by immunization with baculovirus-expressed rickettsia rickettsii rompa protein. | baculovirus recombinants that express the rickettsia rickettsii rompa protein were constructed. monoclonal antibodies (mabs) against the rompa protein reacted with recombinant-infected spodoptera frugiperda (sf9) cells in indirect immunofluorescence assays. sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of infected sf9 cell lysates with a mab against rompa showed that the recombinant-expressed rompa protein migrated slightly below rompa extracted from r. rickettsii. ... | 1995 | 7762273 |