Publications
| Title | Abstract | Year(sorted descending) Filter | PMID Filter |
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| primary infection of insect tracheae by autographa californica m nuclear polyhedrosis virus. | previously we found that autographa californica m nuclear polyhedrosis virus infections initiated in the midguts of insect hosts are disseminated within the hosts via tracheae, elements composing the insect respiratory system. herein we report that infection of tracheae also can be initiated directly via the spiracles, exterior openings of the tracheal system to the atmosphere. infections were initiated both by direct application of viral inoculum to the spiracles and by aerosol. infections esta ... | 1994 | 8030276 |
| expression and post-translational processing of a broad-spectrum organophosphorus-neurotoxin-degrading enzyme in insect tissue culture. | a recombinant baculovirus, autographa californica nuclear polyhedrosis virus (acnpv), has been utilized to express the opd (organophosphate-degrading) gene from pseudomonas diminuta in insect tissue-culture cells (sf9) of the fall armyworm (spodoptera frugiperda). the broad-spectrum organophosphate hydrolase (ec 3.1.8.1) encoded by this gene is a member of a general class of enzymes [organophosphate (op) anhydrolases] that include parathion hydrolases, di-isopropyl-fluorophosphatases (dfpases), ... | 1994 | 8031504 |
| control of programmed cell death by the baculovirus genes p35 and iap. | the sf-21 insect cell line undergoes rapid and widespread apoptosis when treated with actinomycin d or when infected with a mutant of the baculovirus autographa californica nuclear polyhedrosis virus lacking a p35 gene or a functionally active iap (inhibitor of apoptosis) gene. here we provide evidence that the basis for the induction of apoptosis by these two different stimuli is the cessation of rna synthesis. we also show that expression of either p35 or two different functional iap homologs ... | 1994 | 8035800 |
| characterization of glycosylated and catalytically active recombinant human alpha-galactosidase a using a baculovirus vector. | fabry disease is an x-linked inborn error of glycolipid metabolism caused by a deficiency of the lysosomal enzyme alpha-galactosidase a (gala; ec 3.2.1.22). in order to obtain large quantities of this human enzyme for physical characterization and for the development of new approaches for enzyme therapy, we constructed derivatives of the autographa californica nuclear polyhedrosis virus that produce the human enzyme. the recombinant gala (re-gala) is produced at high levels, and is active with b ... | 1994 | 8039705 |
| cloning and expression of bovine and porcine interleukin-2 in baculovirus and analysis of species cross-reactivity. | the cdnas encoding bovine and porcine interleukin-2 (il-2) have been expressed using the baculovirus autographa californica nuclear polyhedrosis virus as a vector in insect cells. insect cells infected with recombinant viruses secreted bovine and porcine il-2 into the culture medium, with biological activities for maintaining the proliferation of homologous cells. when the activities of these two il-2 proteins and commercially available human il-2 were tested on heterologous cells differences we ... | 1994 | 8042283 |
| interaction of trichoplusia ni granulosis virus-encoded enhancin with the midgut epithelium and peritrophic membrane of four lepidopteran insects. | enhancin, an infectivity-enhancing protein from trichoplusia ni granulosis virus (tngv) was tested for its ability to increase autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv) infection in the larvae of four lepidopteran insects. enhancin increased the mortality of acmnpv infection in all the four insect species tested. peritrophic membrane (pm) assays showed altered protein profiles in pms treated with enhancin in all the four species. this supports the hypothesi ... | 1994 | 8046398 |
| the p10 gene of natural isolates of bombyx mori nuclear polyhedrosis virus encodes a truncated protein with an m(r) of 7700. | sequence analysis of the p10 genes of three bombyx mori nuclear polyhedrosis virus (bmnpv) isolates collected in taiwan (ta) and japan (t3 and d1) showed that all possessed a deletion of an adenine residue, 210 bp downstream from the first base of the initiation codon when compared to the p10 gene of autographa californica (multinucleocapsid) npv (acmnpv). this deletion caused a downstream termination codon to come inframe with the coding sequence of p10, so that the p10 gene of bmnpv encoded a ... | 1994 | 8046414 |
| nucleotide sequence of the polyhedrin gene of spodoptera littoralis multiple nucleocapsid nuclear polyhedrosis virus. | 1286 nt portion of the 3.1 kbp hindiii-k fragment of the spodoptera littoralis multiple nucleocapsid nuclear polyhedrosis virus (simnpv) dna genome has been sequenced. the sequence contains the polyhedrin gene preceded by a highly-repeated agataa-rich sequence. with 249 amino acids, simnpv polyhedrin is the longest known polyhedrin. | 1994 | 8049274 |
| direct cloning into the autographa californica nuclear polyhedrosis virus for generation of recombinant baculoviruses. | 1994 | 8052541 | |
| [primary structure of polyhedrin from the nuclear polyhedrosis virus of the silkworm malacosoma neustria]. | 1994 | 8053947 | |
| subcore- and core-like particles of broadhaven virus (brdv), a tick-borne orbivirus, synthesized from baculovirus expressed vp2 and vp7, the major core proteins of brdv. | the genes encoding the two major core proteins (vp2 and vp7) of broadhaven (brd) virus, a tick-borne orbivirus, were inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) under the control of copies of the acnpv polyhedrin promoter to produce two recombinant baculoviruses. infection of spodoptera frugiperda (sf) cells with a recombinant acnpv that synthesized brdv vp2 produced large numbers of brdv subcore-like particles. co-infection of cells with the two recombi ... | 1994 | 8079519 |
| a cysteine protease encoded by the baculovirus bombyx mori nuclear polyhedrosis virus. | sequence analysis of the bamhi f fragment of the genome of bombyx mori nuclear polyhedrosis virus (bmnpv) revealed an open reading frame whose deduced amino acid sequence had homology to those of cysteine proteases of the papain superfamily. the putative cysteine protease sequence (bmnpv-cp) was 323 amino acids long and showed 35% identity to a cysteine proteinase precursor from trypanosoma brucei. of 36 residues conserved among cathepsins b, h, l, and s and papain, 31 were identical in bmnpv-cp ... | 1994 | 8083997 |
| identification of three late expression factor genes within the 33.8- to 43.4-map-unit region of autographa californica nuclear polyhedrosis virus. | a transient transactivation assay system was used in combination with an overlapping autographa californica nuclear polyhedrosis virus clone library to identify genes involved in late and very late baculovirus gene expression. we have identified three genes within the 33.8- to 43.4-map-unit region of the a. californica nuclear polyhedrosis virus genome which contribute to expression from promoters of the vp39 major capsid protein and polyhedrin genes. one of these three genes corresponds to the ... | 1994 | 8084003 |
| influence of infection route on the infectivity of baculovirus mutants lacking the apoptosis-inhibiting gene p35 and the adjacent gene p94. | the infectivity of autographa californica nuclear polyhedrosis virus mutants lacking the apoptosis-inhibiting gene p35 is decreased 1,000-fold or more in larvae of the insect spodoptera frugiperda if the budded form of the virus is administered by hemocoelic injection; this decrease is correlated with the antiviral effects of apoptosis (r. j. clem and l. k. miller, j. virol. 67:3730-3738, 1993). we have extended this correlation by showing that the infectivity of p35 mutant budded virus is resto ... | 1994 | 8084009 |
| transcription, translation, and cellular localization of pdv-e66: a structural protein of the pdv envelope of autographa californica nuclear polyhedrosis virus. | a late gene encoding a 66-kda structural protein of autographa californica nuclear polyhedrosis virus was mapped and sequenced (26.9-29.7 mu). transcription initiates from two conserved taag motifs (-15 and -37) with transcripts detected from 12 to 72 hr pi. the protein is detected in infected spodoptera frugiperda (sf9) cells from 24 to 72 hr pi. western-blot and immunoelectron microscopic data identify this protein as specific for the polyhedra-derived virus (pdv) envelope. this protein has be ... | 1994 | 8091653 |
| genetic analyses of temperature-sensitive mutations in baculovirus late expression factors. | two temperature-sensitive mutants of autographa californica nuclear polyhedrosis virus were characterized with respect to late and very late gene expression. cells infected with ts317 or ts538 at the nonpermissive temperature revealed slightly reduced intracellular levels of late structural gene products including gp64 envelope, vp39 capsid, and p6.9 dna binding proteins. however, with the exception of gp64, there was a dramatic reduction in extracellular levels of the structural proteins, sugge ... | 1994 | 8091663 |
| expression of the 39k promoter of autographa californica nuclear polyhedrosis virus is increased by the apoptotic suppressor p35. | the 39k promoter of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) is transactivated by the viral protein ie1 and further stimulated by viral enhancer elements and the viral coactivator ie2. to identify additional viral proteins that regulate expression from the 39k promoter, we performed cotransfection experiments with clones of viral dna and a plasmid containing the chloramphenicol acetyltransferase (cat) gene under the control of the 39k promoter. our results indica ... | 1994 | 8091669 |
| 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 |
| expression of semliki forest virus nsp1-specific methyltransferase in insect cells and in escherichia coli. | we have expressed the semliki forest virus (sfv)-specific nonstructural protein nsp1 both in insect cells and in escherichia coli in the absence of other viral proteins. a substantial amount of nsp1 was synthesized in sf9 cells infected with the recombinant autographa californica nuclear polyhedrosis virus (acnpv) acnpv-nsp1. these cells had a high level of guanine-7-methyltransferase activity compared with that of wild-type acnpv-infected cells. the methyltransferase activity and nsp1 were most ... | 1994 | 7933125 |
| 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 |
| overexpression of human dna topoisomerase i in insect cells using a baculovirus vector. | the 3645-bp human dna topoisomerase i cdna isolated by d'arpa et al. (proc. natl. acad. sci. usa 85, 1988, 2543-2547) was integrated into the autographa californica multiple nuclear polyhedrosis virus genome. the recombinant protein was expressed by infecting the sf9 insect cell line with this baculovirus and resulted in a 100-fold overexpression of human dna topoisomerase i compared to the level found in human cell lines. this 100-kda recombinant protein has the same electrophoretic mobility as ... | 1994 | 7950383 |
| overexpression, purification, and characterization of third component of complement. | the third component of complement (c3) plays a critical role in both pathways of complement activation by interacting with numerous other complement proteins. to elucidate the molecular features of c3 that relate to the functional activities of the molecule, we expressed the cdna of human complement component c3 in cultured insect cells using a baculovirus expression vector system derived from the baculovirus autographa california nuclear polyhedrosis virus (acnpv). the expression of c3 was cont ... | 1994 | 7963591 |
| the pk-1 gene of autographa californica multinucleocapsid nuclear polyhedrosis virus encodes a protein kinase. | open reading frame (orf) 9 of the ecori i fragment of the baculovirus autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) genome encodes a protein, pk-1, which has strong similarity to serine-threonine protein kinases. the published sequence of the pk-1 gene contains an error that, when corrected, extends the orf by 228 nucleotides. transcription of pk-1 was first detectable by primer extension at 12 h post-infection, accumulating to high levels during the very late phas ... | 1994 | 7964609 |
| nucleotide sequence of the polyhedrin gene region of helicoverpa zea single nucleocapsid nuclear polyhedrosis virus: placement of the virus in lepidopteran nuclear polyhedrosis virus group ii. | the polyhedrin gene (polh) of helicoverpa zea single nucleocapsid nuclear polyhedrosis virus (hzsnpv) was identified and shown by sequence analysis of the ecori i genomic fragment to encode a 246 amino acid polypeptide that has greater than 80% sequence identity to known polyhedrins. it is preceded by an at-rich region containing the conserved late promoter motif taag, which was identified as a transcription start point. downstream of polh there were several similarities in genome arrangement to ... | 1994 | 7964631 |
| 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 |
| identification and characterization of vlf-1, a baculovirus gene involved in very late gene expression. | we have identified a gene required for strong expression of the polyhedrin gene by characterizing a mutant, tsb837, of the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) which is temperature sensitive (ts) for occluded virus production at the nonpermissive temperature. marker rescue experiments utilizing an overlapping set of acmnpv genomic clones revealed that the gene responsible for the ts mutant phenotype mapped to a region between 46 and 48 map units. fragments (2.2 ... | 1994 | 7966564 |
| biosynthesis and processing of the autographa californica nuclear polyhedrosis virus gp64 protein. | gp64 is a major virion envelope glycoprotein of the baculovirus autographa californica multicapsid nuclear polyhedrosis virus (acmnpv). gp64 plays an important role in acmnpv infection, probably mediating penetration of one form of the virus into host cells through the endocytic pathway. gp64 also represents an excellent probe for studying the membrane glycoprotein processing capabilities of baculovirus-infected insect cells, which are used widely as a eucaryotic expression system. the goals of ... | 1994 | 7975226 |
| expression of recombinant rabbit udp-glcnac: alpha 3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i catalytic domain in sf9 insect cells. | udp-glcnac: alpha 3-d-mannoside beta-1,2-n-acetylglucosaminyltransferase i (gnt i; ec 2.4.1.101) catalyses a key reaction in the conversion of oligomannose to complex and hybrid n-glycans. the cytoplasmic tail and transmembrane segment of rabbit gnt i cdna were replaced with an in-frame cleavable signal sequence and the hybrid construct was inserted into the genome of autographa californica nuclear polyhedrosis virus (acmnpv) under the control of the polyhedrin promoter. sf9 insect cells were in ... | 1994 | 7841795 |
| 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 |
| recombination between genetically modified and unmodified autographa californica nuclear polyhedrosis virus in trichoplusia ni larvae. | trichoplusia ni larvae have been injected with a mixture of wild-type autographa californica multiple nuclear polyhedrosis virus (acmnpv) and a mutant derivative, acrp8.uw1.lacz, which lacks the polyhedrin gene, and has the p10 gene replaced by the escherichia coli beta-galactosidase gene. following plaque assay of the haemolymph and subsequent staining for beta-galactosidase activity and scoring for polyhedra, recombinant plaques were identified and the recombination frequency estimated as 6.6% ... | 1994 | 7793354 |
| 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 |
| evidence for the ligand-induced conversion from a dimer to a tetramer of the granulocyte colony-stimulating factor receptor. | an extracellular portion of granulocyte colony-stimulating factor (g-csf) receptor, which contains an immunoglobulin-like (ig) domain and cytokine receptor homologous (crh) region, was secreted into the medium using trichoplusia ni-autographa californica nuclear polyhedrosis virus system. the gene product was purified to homogeneity mainly as a dimer (85 kda) using g-csf affinity column chromatography and gel filtration hplc, although the product existed as a monomer (45 kda) in the medium. scat ... | 1994 | 7805849 |
| nuclear export of recombinant baculovirus nucleocapsids through small pore or nuclear-pore-like structure in sf9 cells. | translocation of baculovirus nucleocapsids (45 nm in diameter, approximate length of 280-300 nm) from nucleoplasm to cytoplasm was studied morphologically using cryofixation and gold labeled wheat germ agglutinin (wga-gold) during recombinant autographa californica nuclear polyhedrosis virus infection in sf9 cells. nucleocapsids formed in the nucleoplasm migrated into protrusions of the nuclear envelope, but not into nuclear pore complexes. we found cross-like membranous structures. small pores ... | 1994 | 7808726 |
| uses of flow cytometry in virology. | this article reviews some of the published applications of flow cytometry for in vitro and in vivo detection and enumeration of virus-infected cells. sample preparation, fixation, and permeabilization techniques for a number of virus-cell systems are evaluated. the use of flow cytometry for multiparameter analysis of virus-cell interactions for simian virus 40, herpes simplex viruses, human cytomegalovirus, and human immunodeficiency virus and its use for determining the effect of antiviral comp ... | 1994 | 7530594 |
| 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 |
| a putative non-hr origin of dna replication in the hindiii-k fragment of autographa californica multiple nucleocapsid nuclear polyhedrosis virus. | in addition to the seven known homologous regions (hrs) of autographa californica multiple nucleocapsid polyhedrosis virus (acmnpv) the hindiii-k fragment was also found to carry a putative ori, although this fragment does not contain an hr. deletion analysis showed that this ori contains several segments essential for its activity and other 'auxiliary' sequences that enhance the ori activity. sequence analysis identified several structures often found in other viral replication oris, such as pa ... | 1994 | 7996129 |
| the granulin gene region of cryptophlebia leucotreta granulosis virus: sequence analysis and phylogenetic considerations. | the nucleotide sequence of a 2.2 kb genome region of the baculovirus cryptophlebia leucotreta granulosis virus (cigv) encompassing the granulin gene and a second open reading frame, designated orf909, was determined. the putative granulin orf comprises 744 nucleotides encoding a polypeptide with a predicted m(r) of 29.3 k. the 5' leader of the granulin gene contains a canonical late baculovirus promoter (ataag) and differs only in two nucleotide positions from the sequence of cydia pomonella gra ... | 1994 | 7996162 |
| 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 |
| 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 |
| quantification of latent mamestra brassicae nuclear polyhedrosis virus in m. brassicae insects using a pcr-scintillation proximity assay. | a laboratory culture of mamestra brassicae insects (mblc) was found to harbour a latent baculovirus infection. the copy number of the occult mbnpv genome in both the mblc larvae, and in a cell line derived from the fat body of mblc was determined by the use of a rapid and convenient pcr-scintillation proximity assay (spa). the spa system relies on the use of fluomicrospheres (spa beads) coated with acceptor molecules which are capable of binding radiolabelled ligands in solution. in the assay de ... | 1994 | 7714043 |
| identification and characterization of a protein kinase gene in the lymantria dispar multinucleocapsid nuclear polyhedrosis virus. | the lymantria dispar multinucleocapsid nuclear polyhedrosis virus (ldmnpv) gene encoding vpk has been cloned and sequenced. ldmnpv vpk shows a 24% amino acid identity to the catalytic domains of the eucaryotic protein kinases npkc from rabbits, hspkce from humans, aplpkcb from aplysia californica, and dpkc98f from drosophila melanogaster, and homology to several other protein kinases from yeasts, mice, and bovines. the homology suggests that vpk is a serine/threonine protein kinase as defined by ... | 1994 | 8107234 |
| nucleotide sequence and genetic organization of a 7.3 kb region (map unit 47 to 52.5) of autographa californica nuclear polyhedrosis virus fragment ecori-c. | the nucleotide sequence and genetic organization of a 7297 bp region within the ecori-c fragment of autographa californica multiple nucleocapsid nuclear polyhedrosis virus (acmnpv) are presented. eight putative open reading frames were found and their respective amino acid sequences compared with a number of data libraries. orf 1227 corresponded with gp41 and its predicted protein sequence was found to be 55 amino acids longer at its c terminus than reported previously. moreover the main part of ... | 1994 | 8126447 |
| deletion and mutational analyses of bluetongue virus ns2 protein indicate that the amino but not the carboxy terminus of the protein is critical for rna-protein interactions. | genome segment 8 (s8) of bluetongue virus serotype 10 (btv-10) encodes the nonstructural protein ns2. this protein, which has single-stranded rna (ssrna) binding capacity, is found in btv-infected cells in the form of virus inclusion bodies (vibs). to identify the domain(s) important for rna binding and oligomerization of the protein, a number of deletions were made in regions of the gene that code for either the amino or carboxy terminus of the protein. the modified genes were cloned into and e ... | 1994 | 8139002 |
| an apoptosis-inhibiting gene from a nuclear polyhedrosis virus encoding a polypeptide with cys/his sequence motifs. | two different baculovirus genes are known to be able to block apoptosis triggered upon infection of spodoptera frugiperda cells with p35 mutants of the insect baculovirus autographa californica nuclear polyhedrosis virus (acmnpv):p35 (p35-encoding gene) of acmnpv (r. j. clem, m. fechheimer, and l. k. miller, science 254:1388-1390, 1991) and iap (inhibitor of apoptosis gene) of cydia pomonella granulosis virus (cpgv) (n. e. crook, r. j. clem, and l. k. miller, j. virol. 67:2168-2174, 1993). using ... | 1994 | 8139034 |
| temporal regulation of a complex and unconventional promoter by viral products. | the dna polymerase (dnapol) gene of autographa californica nuclear polyhedrosis virus presents a complex promoter organization. it lacks the usual tata box and start site, and its rna accumulation initially increases and then decreases dramatically during infection. we investigated dnapol temporal regulation. transiently expressed dnapol gene was transcribed at a low level from minor start sites. coexpression with ie0 and/or ie1 immediate-early genes dramatically enhanced dnapol transcription, s ... | 1994 | 8139038 |
| mutational analysis of a baculovirus major late promoter. | we have used a linker-scan mutation strategy to analyze pcap99, the proximal promoter of the autographa californica nuclear polyhedrosis virus (acmnpv) gene encoding the major capsid protein. a series of recombinant viruses expressing the cat reporter gene under the control of selected mutants of this promoter was constructed. only mutations that altered bases within a region extending from 8 bp upstream to 6 bp downstream from a taag sequence had a significant effect on expression from the late ... | 1994 | 8144021 |
| human monocyte chemoattractant protein-1 expressed in a baculovirus system. | human monocyte chemoattractant protein-1 (hmcp-1) was produced using a baculovirus system. the hmcp-1 cdna was inserted into the genomic dna of autographa californica nuclear polyhedrosis virus (acnpv) using a transfer vector, pjvp10z. spodoptera frugiperda insect cells, which were infected with this recombinant virus, secreted recombinant hmcp-1 (re-hmcp-1) at the level of 10-20 micrograms/ml of culture medium. this product was shown to chemoattract monocytes. three distinct bands of 11, 11.5 a ... | 1994 | 8144036 |
| dengue type-2 virus envelope protein made using recombinant baculovirus protects mice against virus challenge. | the gene coding for the envelope (e) glycoprotein of dengue-2 virus was cloned into baculovirus (autographa californica nuclear polyhedrosis virus). the recombinant virus contained the entire e protein gene, preceded by 38 nucleotides from the end of the prematrix glycoprotein gene and followed by the first 83 nucleotides of nonstructural protein 1. when expressed in spodoptera frugiperda (sf9) cells, approximately 1 mg of recombinant e antigen was made per 10(9) cells. this antigen reacted with ... | 1994 | 8147490 |
| identification and characterization of the ecdysteroid udp-glucosyltransferase gene of the lymantria dispar multinucleocapsid nuclear polyhedrosis virus. | we have located, cloned, sequenced and characterized the ecdysteroid udp-glycosyltransferase gene (egt) gene from the baculovirus lymantria dispar multinucleocapsid nuclear polyhedrosis virus. (ldmnpv), which is specific for the gypsy moth (l. dispar). the egt gene from the related baculovirus autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) disrupts the hormonal balance of the host larva by galactosylating ecdysone, which prevents moulting. the location of the ldmnpv ... | 1994 | 8151299 |
| the insect tracheal system: a conduit for the systemic spread of autographa californica m nuclear polyhedrosis virus. | baculoviruses establish systemic infections within susceptible insect hosts, even though host tissues are surrounded by basal laminae, extracellular matrices that exclude particles smaller than these viruses. using a recombinant autographa californica m nuclear polyhedrosis virus containing a lacz reporter gene under the control of a constitutive promoter, we followed the progression of infection in trichoplusia ni larvae. we discovered that infection of the larval insect tracheal system (and no ... | 1994 | 8159729 |
| functional dissection of the autographa california nuclear polyhedrosis virus enhancer element hr5. | the hr5 enhancer element of autographa californica nuclear polyhedrosis virus stimulates transcription from baculovirus-delayed early promoters in the presence of the viral transactivator, ie1. gel retardation analyses of interactions between a fragment of hr5 and extracts prepared from cells transfected with an ie1-encoding plasmid revealed the presence of three dna-protein complexes. in order to better define the functional domains of the hr5 enhancer, we constructed a set of plasmids containi ... | 1994 | 8178424 |
| identification of lef-7: a baculovirus gene affecting late gene expression. | we have examined a 15-kb region of the autographa californica nuclear polyhedrosis virus (acmnpv) genome, from 66 to 78 map units, for the presence of genes which transactivate expression from late and very late viral promoters in transient expression assays. one gene in this region activated reporter gene expression approximately six- to eightfold when the reporter gene was under the control of the late promoter of the major capsid protein gene, vp39, or the very late promoter of the polyhedrin ... | 1994 | 8178427 |
| specificity of baculovirus p10 functions. | three functional domains in baculovirus p10 proteins have been postulated for aggregation, nuclear disintegration, and fibrillar structure formation (van oers et al., j. gen. virol. 74, 563-574, 1993). to study the specificity of these functions, a recombinant autographa californica nuclear polyhedrosis virus (accr1) was constructed in which the coding sequence of the p10 gene was replaced with the p10 sequence of the distantly related spodoptera exigua (se) mnpv. in accr1-infected cells the sem ... | 1994 | 8178439 |
| synthesis of recombinant baculoviruses expressing the outer capsid protein vp2 of five btv serotypes and the induction of neutralizing antibodies to homologous and heterologous btv serotypes. | dna representing rna segment l2 of 5 different bluetongue virus (btv) serotypes (btv-1, -2, -11, -13 and -17) corresponding to the gene that codes for the btv neutralization antigen vp2, have been inserted individually into baculovirus transfer vectors in lieu of the 5' coding region of the polyhedrin gene of autographa california nuclear polyhedrosis virus (acnpv). after co-transfection of spodoptera frugiperda cells with wild-type acnpv dna in the presence of the derived transfer vector dnas, ... | 1994 | 8178571 |
| [physical mapping of the bombyx mori nuclear polyhedrosis virus genome]. | the restriction map of the nuclear polyhedrosis dna virus of bombyx mori has been determined with the use of four restriction endonucleases: noti, bamhi, smai and kpni. 18 cleavage sites have been localized on the viral dna. total length of the viral dna is about 130 kb. the fragment containing a polyhedrin gene has been localized on the map by the blot hybridization technique. | 1994 | 8183306 |
| secretion of recombinant rat annexin 5 by insect cells in a baculovirus expression system. | rat annexin 5 was expressed in insect cells using a baculovirus vector, autographa californica nuclear polyhedrosis virus. the rat annexin 5 cdna was prepared by a polymerase chain reaction using mrna from rat pituitary glands and placed under the control of the polyhedrin promoter. the gene product was 36 k dalton and was recognized by anti-rat annexin 5 serum. the calcium dependent binding of the recombinant annexin 5 to membranes was confirmed. the recombinant protein appeared in the medium b ... | 1994 | 8185595 |
| flow cytometric analysis of bioluminescence emitted by recombinant baculovirus-infected insect cells. | five recombinant baculoviruses, each containing a different insect luciferase gene encoding a protein with characteristic light emission properties, namely, luc gr (546 nm), luc ff (556 nm), luc yg (560 nm), luc ye (578 nm), and luc or (593 nm) were constructed. all genes were inserted under the transcriptional control of the polyhedrin gene promoter of the autographa californica nuclear polyhedrosis virus (acnpv) and expressed in spodoptera frugiperda insect cells during viral infection. the bi ... | 1994 | 8187580 |
| the apoptotic suppressor p35 is required early during baculovirus replication and is targeted to the cytosol of infected cells. | the p35 gene of autographa californica nuclear polyhedrosis virus (acmnpv) is required to block virus-induced apoptosis. the trans-dominant activity of p35 suppresses premature cell death and facilitates acmnpv replication in a cell line- and host-specific manner. to characterize the p35 gene product (p35), a specific polyclonal antiserum was raised. as revealed by immunoblot analyses of wild-type acmnpv-infected cells, p35 appeared early (8 to 12 h) and accumulated through the late stages of in ... | 1994 | 8189486 |
| detection of a putative 30-kda ligand of the cluster-2 antigen. | the cluster-2 antigen, also called egp-2, is a 38-kda transmembrane glycoprotein with a distribution that is largely confined to human epithelial cells and their derived carcinomas. monoclonal antibodies (mabs) directed against egp-2 have been extensively studied as anti-tumor agents, yet the function of the antigen is not known. in the present study we used a biotinylated recombinant soluble derivative of the egp-2 (segpbio) as a probe to detect a possible egp-2 ligand, using various carcinoma ... | 1994 | 8194899 |
| expression and characterization of human bone morphogenetic protein-2 in silkworm larvae infected with recombinant bombyx mori nuclear polyhedrosis virus. | recombinant human bone morphogenetic protein-2 (rhbmp-2) was expressed in silkworm larvae, and a milligram quantity of the protein was purified and characterized. the expressed rhbmp-2 was biologically active in terms of induction of alkaline phosphatase activity in mc3t3-e1 cells and ectopic bone formation in mice. on sds-polyacrylamide gel electrophoretic analysis, the purified protein showed a 16 kda band under reducing conditions and a 30 kda band under non-reducing conditions. the silkworm- ... | 1994 | 8206877 |
| mutagenesis of recombinant protein c inhibitor reactive site residues alters target proteinase specificity. | protein c inhibitor (pci) is a heparin-binding plasma serine proteinase inhibitor (serpin) which is thought to be a physiological regulator of activated protein c. we are using recombinant pci (rpci) to study structural determinants of target proteinase specificity. a cdna encoding full-length pci has been expressed as a fully active proteinase inhibitor using autographa californica nuclear polyhedrosis virus (baculovirus). rpci was expressed maximally 4 days after infection and could be express ... | 1994 | 8206990 |
| identification and transcriptional regulation of the baculovirus lef-6 gene. | we identified an autographa californica nuclear polyhedrosis virus (acmnpv) gene, the late expression factor 6 gene (lef-6), which is involved in expression from late and very late acmnpv gene promoters but not from an early acmnpv gene promoter in transient expression assays. this gene was located within the psti i fragment of the acmnpv genome (14.7 to 17.9 map units), immediately downstream of ac-iap, the acmnpv homolog of a baculovirus gene family involved in blocking apoptotic programmed ce ... | 1994 | 8207819 |
| a baculovirus gene involved in late gene expression predicts a large polypeptide with a conserved motif of rna polymerases. | we have identified and sequenced a novel baculovirus gene, late expression factor eight gene (lef-8), of autographa californica nuclear polyhedrosis virus that is necessary for efficient expression from late and very late virus gene promoters in a transient expression assay. the predicted gene product, lef-8, has a molecular mass of 102 kda and contains a conserved sequence motif, gxkx4hgq/nkg, found in dna-directed rna polymerases throughout the animal, plant, and microbial kingdoms. | 1994 | 8207843 |
| an unusual 30-kda protein binding to the polyhedrin gene promoter of autographa californica nuclear polyhedrosis virus. | gel retardation assays using nuclear extracts from the autographa californica nuclear polyhedrosis virus-infected sf21 (spodoptera frugiperda) insect cell line revealed a host factor (polyhedrin promoter-binding protein or ppbp) that binds to the polyhedrin gene promoter. a hexanucleotide sequence (aataaa) within the promoter is important for binding in association with neighboring elements, some of which are contributed by the sequence taagtatt present at the transcription start point. ppbp was ... | 1994 | 8300607 |
| extension of autographa californica nuclear polyhedrosis virus host range by interspecific replacement of a short dna sequence in the p143 helicase gene. | recombinant baculoviruses obtained by coinfection of insect cells with autographa californica and bombyx mori nuclear polyhedrosis viruses (acnpv and bmnpv, respectively) possess a wider in vitro host range than either parent virus. to localize the dna sequences responsible for this species specificity, we used a two-step method of production and selection of recombinant viruses with altered specificity. sf9 cells, which are permissive for acnpv, were first cotransfected with genomic acnpv dna a ... | 1994 | 8278405 |
| a mutant baculovirus with a temperature-sensitive ie-1 transregulatory protein. | we have mapped the mutation responsible for the temperature-sensitive (ts) phenotype of tsb821, a mutant of the baculovirus autographa californica nuclear polyhedrosis virus (h. h. lee and l. k. miller, j. virol. 31:240-252, 1979), to a single nucleotide which changes alanine 432 of the multifunctional regulatory protein ie-1 to a valine. mapping was done with a combination of marker rescue and transient expression assays, hybrid gene construction by overlap pcr gene splicing, and nucleotide seq ... | 1994 | 8289337 |
| in vivo and in vitro analyses of recombinant baculoviruses lacking a functional cg30 gene. | the cg30 gene of autographa californica nuclear polyhedrosis virus (acmnpv) encodes two sequence motifs, a zinc finger-like motif and a leucine zipper, found in other polypeptides known to be involved in gene regulation. to gain insight into the function of the cg30 product, cg30, we constructed and characterized recombinant viruses lacking a functional cg30 gene. we found that cg30 mutants had no striking phenotype in cell lines derived from spodoptera frugiperda or trichoplusia ni or in t. ni ... | 1994 | 8289348 |
| high-level expression and purification of secreted forms of herpes simplex virus type 1 glycoprotein gd synthesized by baculovirus-infected insect cells. | two forms of herpes simplex virus glycoprotein gd were recombined into autographa californica nuclear polyhedrosis virus (baculovirus) and expressed in infected spodoptera frugiperda (sf9) cells. each protein was truncated at residue 306 of mature gd. one form, gd-1(306t), contains the coding sequence of patton strain herpes simplex virus type 1 gd; the other, gd-1(qaat), contains three mutations which eliminate all signals for addition of n-linked oligosaccharides. prior to recombination, each ... | 1994 | 8289380 |
| a baculovirus gp64 early promoter is activated by host transcription factor binding to cacgtg and gata elements. | the early promoter of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus gp64 gene is active when transfected into several insect cell lines and does not require viral gene products for transcription in uninfected cells. because previous studies have shown that the gp64 early promoter is activated above basal levels in uninfected cells, host transcription factors are likely to play a role in gp64 activation at early times postinfection. by using nuclear extracts from uninfected sf9 ... | 1994 | 8289385 |
| functional mapping of regions of the autographa californica nuclear polyhedrosis viral genome required for dna replication. | previous results showed that plasmids containing one of the eight putative origins (ori's) of autographa californica nuclear polyhedrosis virus (acmnpv) are replicated after transfection into spodoptera frugiperda cells if essential trans-acting factors are supplied by acmnpv infection (kool et al., virology, 192, 94-101, 1993a; kool et al., j. gen. virol., in press, 1993b; leisy and rohrmann, virology, 196, 722-730, 1993). in this report a transient complementation assay is described in which f ... | 1994 | 8291249 |
| glycosylation and high-level secretion of human tumour necrosis factor-beta in recombinant baculovirus-infected insect cells. | human tumour necrosis factor-beta (tnf-beta) was produced in eukaryotic cells using the insect baculovirus cloning and expression system. a novel insect signal sequence, the honey-bee (apis mellifera) prepromelittin secretory sequence, was used to aid in the post-translational modifications, glycosylation and secretion of recombinant human tnf-beta. human tnf-beta cdna was cloned using the insect baculovirus vector pacc4s. expression of the human tnf-beta was regulated by the insect autographa c ... | 1993 | 8297505 |
| the 1629-bp open reading frame of the autographa californica multinucleocapsid nuclear polyhedrosis virus encodes a virion structural protein. | a 1629-bp open reading frame (orf) of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) is shown to encode a 78-kda virion structural protein. to determine this, polyclonal antibody was made to a fusion protein synthesized in escherichia coli from a chimeric gene that contained 1415 bp of the 1629-bp gene. in western blot analyses, this antibody cross-reacted with a protein of about 78 kda in both extracellular virions (ecv) and virions isolated from polyhedra (pdv), a ... | 1993 | 8299959 |
| baculovirus-directed expression of human prostatic steroid 5 alpha-reductase 1 in an active form. | in the human prostate, the enzyme steroid 5 alpha-reductase (h5 alpha r) catalyses the conversion of testosterone into the more potent androgen, dihydrotestosterone. two distinct cdnas coding for h 5 alpha r in the human prostate have been previously characterized. enzyme h5 alpha r1 shows a maximum activity at basic ph whereas h5 alpha r2 has an acidic ph optimum activity. we report here the expression of the human steroid h5 alpha r1 in a eukaryotic expression system: the baculovirus-directed- ... | 1993 | 8664165 |
| identification and immunocharacterization of a 25k structural protein in autographa californica nuclear polyhedrosis virus (acmnpv). | a 25k protein was a major component of the triton x-100 solubilized fraction prepared for the isolation of nuclear matrices from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acmnpv). polyclonal antisera to p25 were produced in rabbits and used in the characterization of p25. western blots of proteins probed with the p25 antiserum demonstrated the presence of the p25 in both the polyhedral-derived and extracellular viral phenotypes. using an indirec ... | 1993 | 8317143 |
| non-homologous recombination between adenovirus and acnpv dna fragments in cell-free extracts from insect spodoptera frugiperda nuclei. | in previous work, we have developed a cell-free system from nuclear extracts of hamster cells to study the mechanism of integrative recombination between adenovirus type 12 (ad12) dna and hamster cell dna (jessberger et al., 1989; tatzelt et al., 1992). we have also demonstrated that in insect cells the left terminal fragment of ad2 dna can insert by non-homologous recombination into the 32.6 to 34.5 map unit (ecori-o) fragment and into other segments of autographa californica nuclear polyhedros ... | 1993 | 8317144 |
| one-step purification and characterization of human pancreatic lipase expressed in insect cells. | a cdna clone encoding the sequence of human pancreatic lipase (hpl) was subcloned into the baculovirus transfer vector pvl1392 and used in co-transfection of spodoptera frugiperda (sf9) insect cells with wild-type autographa californica nuclear polyhedrosis virus (acnpv) dna. a single recombinant protein (50 kda) secreted by sf9 cells was detectable in the culture medium 24 h post-infection using both anti-hpl polyclonal antibodies and potentiometric measurements of lipolytic activity. the expre ... | 1993 | 8335100 |
| a 25-kda protein is associated with the envelopes of occluded baculovirus virions. | antiserum produced against preoccluded virions of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was used to screen an opmnpv lambda gt11 expression library. one of the immunoreactive clones contained an insert that hybridized to a portion of the opmnpv hindiii-p fragment. a 2-kb region of hindiii-p was sequenced and found to contain three open reading frames, each preceded by a late promoter element. the insert from the lambda gt11 clone was derived from the thir ... | 1993 | 8337828 |
| transcription mapping and functional analysis of the protein tyrosine/serine phosphatase (ptpase) gene of the autographa californica nuclear polyhedrosis virus. | the protein tyrosine/serine phosphatase (ptpase) gene of the autographa californica nuclear polyhedrosis virus yields two major transcripts of approximate sizes 3.1 and 3.9 kb. both of these are very late transcripts, which accumulate to maximal levels more than 30 hr postinfection. the smaller transcript initiates at the t of an ataag sequence that lies 22 bp upstream of the putative start codon of the gene. the larger transcript initiates at the first a of a ttaag sequence that lies approximat ... | 1993 | 8337833 |
| immediate-early baculovirus genes transactivate the p143 gene promoter of autographa californica nuclear polyhedrosis virus. | trans-acting regulatory components of autographa californica nuclear polyhedrosis virus (acmnpv) were studied in a transient assay system for their ability to activate the p143 gene promoter. a region including 502 nt upstream of the aug of the p143 gene was linked to the bacterial chloramphenicol acetyltransferase (cat) gene. the p143 promoter-cat construct was used to identify acmnpv immediate-early gene products required for expression from the p143 gene early promoter. transient expression a ... | 1993 | 8337840 |
| characterization of an early gene coding for a highly basic 8.9k protein from the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. | a new gene of the baculovirus orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) has been identified that encodes a highly basic 8.9k protein. the gene called p8.9 is expressed as a 0.5 kb transcript by 1 h post-infection and initiates at an early gene motif. the promoter of the 0.5 kb transcript has two upstream elements, repeats i and ii, which are similar to motifs previously characterized in the opmnpv ie-2 gene and the autographa californica nuclear polyhedrosis virus ie-n ... | 1993 | 8345350 |
| nucleotide sequence of the buzura suppressaria single nucleocapsid nuclear polyhedrosis virus polyhedrin gene. | a portion of the genome of the buzura suppressaria (lepidoptera) single nucleocapsid nuclear polyhedrosis virus (bssnpv) containing the polyhedrin gene was sequenced. an open reading frame of 738 nucleotides encoded a protein of 246 amino acids and represented the polyhedrin gene. a conserved taag motif, associated with transcriptional start sites in other polyhedrin genes, was identified 51 nucleotides upstream of the bssnpv polyhedrin gene. a putative polyadenylation signal, aataaa, was found ... | 1993 | 8345353 |
| expression and characterization of biologically active human hepatocyte growth factor (hgf) by insect cells infected with hgf-recombinant baculovirus. | a cdna containing the entire coding sequence of human hepatocyte growth factor (hgf) [also known as scatter factor (sf)] was inserted into the genome of autographa california nuclear polyhedrosis virus (baculovirus) adjacent to the polyhedrin promoter by homologous recombination. insect cells (spodoptera frugiperda) infected with the recombinant virus secrete relatively high levels (3-8 mg/l) of biologically active hgf into the culture medium. the recombinant hgf induces pronounced morphological ... | 1993 | 8347597 |
| identification, sequence, and transcriptional mapping of lef-3, a baculovirus gene involved in late and very late gene expression. | a trans-acting gene required for late viral gene expression in transient expression assays was identified in the genome of autographa californica nuclear polyhedrosis virus. a genomic library of a. californica nuclear polyhedrosis virus dna lacking a clone spanning the region from 43 to 48 map units (mu) was unable to activate gene expression from a reporter plasmid when the reporter gene was driven by a baculovirus late or very late promoter in transient expression assays. the genomic region re ... | 1993 | 8350397 |
| expression of human il-2 receptor alpha- and beta-chains using the baculovirus expression system. | the genes encoding the alpha- and beta-chains of the human interleukin-2 receptor were expressed in lepidopteran insect cells using the baculovirus expression vector system. the corresponding genes were inserted under the polyhedrin promoter of the autographa california nuclear polyhedrosis virus and expressed in the spodoptera frugiperda insect cell line during viral infection. the recombinant receptor proteins were identified in the insect cell lysates by using protein dot blot and elisa techn ... | 1993 | 8356402 |
| expression in baculovirus vector system of the nucleocapsid protein gene of rinderpest virus. | the rinderpest (rv) nucleocapsid (np) gene segment was inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) adjacent to the polyhedrin promoter. the expression of np protein in sf9 cells was confirmed by indirect immunofluorescence and by western blotting analysis with monoclonal antibodies. recombinant rv-np protein was purified by ultracentrifugation on a sucrose density gradient, and used as an antigen for an enzyme linked immunosorbent assay to detect anti rv ... | 1993 | 8366167 |
| the hr5 transcriptional enhancer stimulates early expression from the autographa californica nuclear polyhedrosis virus genome but is not required for virus replication. | autographa californica nuclear polyhedrosis virus (acmnpv) contains five homologous regions (hr1 through hr5) interspersed throughout its genome. analysis of plasmid transfections indicates that the hrs function as transcriptional enhancers and possible origins of viral dna replication. the role of these repetitive elements in regulating expression from the acmnpv genome was examined by constructing a series of recombinant viruses that tested the effect of hr5 on expression of the adjacent p35 g ... | 1993 | 8371342 |
| alternative transcriptional initiation as a novel mechanism for regulating expression of a baculovirus trans activator. | in this report, we show that the orgyia pseudotsugata nuclear polyhedrosis virus p34 gene, which is homologous to the autographa californica nuclear polyhedrosis virus pe-38 gene, is a trans activator. the predicted p34 protein contains a number of motifs that are similar to those found in other eukaryotic transcriptional trans activators, including a putative zinc finger dna-binding domain, a glutamine-rich domain, and a leucine zipper. northern (rna) blot analysis showed that the p34 gene is e ... | 1993 | 8371344 |
| the 1,629-nucleotide open reading frame located downstream of the autographa californica nuclear polyhedrosis virus polyhedrin gene encodes a nucleocapsid-associated phosphoprotein. | a 78-kda protein was produced in bacteria from a clone of the 1,629-nucleotide open reading frame located immediately downstream from the polyhedrin gene of autographa californica nuclear polyhedrosis virus. the identity of this protein was confirmed by its reactivity with peptide antiserum and amino terminal peptide sequencing after purification from transformed bacteria. the polypeptide was used to produce polyclonal antisera in rabbits. immunoblot analysis of insect cells infected with the ba ... | 1993 | 8371345 |
| sequence and temporal appearance of the early transcribed baculovirus gene he65. | we have identified the early transcribed he65 gene by screening a cdna library from polyadenylated rna which was isolated at 1 h after infection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus (acnpv). nucleotide sequencing analysis of the he65-specific cdna clone reveals one open reading frame of 1,662 nucleotides from which a protein of 65 kda in size can be predicted. the he65 gene is located downstream of the late transcribed p80 gene and upstream of the ... | 1993 | 8371346 |
| characterization of the replication of plasmids containing hr sequences in baculovirus-infected spodoptera frugiperda cells. | the replication of a series of plasmids, each containing one or more of 6 homologous regions (hrs) from the autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) genome, was investigated in acmnpv-infected spodoptera frugiperda cells. although a construct containing hr2 showed the most efficient replication, the sequences flanking the hr region appeared to influence the levels of replication. plasmids with non-hr-containing acmnpv inserts showed greatly reduced replication ... | 1993 | 8372444 |
| adenine dna methyltransferase m.cviri expression accelerates apoptosis in baculovirus-infected insect cells. | the adenine dna methyltransferase m.cviri (tgcma) gene from chlorella virus xz-6e was cloned into the autographa californica nuclear polyhedrosis virus (acmnpv) genome and expressed in spodoptera frugiperda insect cells under the control of two tandemly arranged viral promoters, the early etl promoter and the late polyhedrin promoter. m.cviri activity was first detected at 10 hr p.i and reached a maximum at 48 hr p.i. viral dna synthesized in insect cells infected with m.cviri expressing virus ( ... | 1993 | 8372450 |
| a gene encoding a highly expressed spindle body protein of heliothis armigera entomopoxvirus. | the gene encoding the most abundant protein of purified preparations of heliothis armigera entomopoxvirus (haepv) has been cloned and sequenced. the gene sequence encodes a 40.1k polypeptide with a putative n-terminal 20 amino acid leader peptide, and a single potential n-glycosylation site. analysis of the protein, which has an apparent m(r) of 50k on polyacrylamide gels, confirmed post-translational loss of the leader peptide, but showed no evidence of glycosylation. the protein is related to ... | 1993 | 8376960 |
| analysis of the orgyia pseudotsugata multicapsid nuclear polyhedrosis virus trans-activators ie-1 and ie-2 using monoclonal antibodies. | we have produced monoclonal antibodies against two orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (opmnpv) transcriptional trans-activators, ie-1 and ie-2. temporal analysis of ie-1 and ie-2 proteins have shown that ie-1 continues to increase in steady-state levels from 0 to 120 h post-infection, whereas ie-2 declines by 36 h post-infection. at least five different electrophoretic forms of ie-1 are present in opmnpv-infected ld652y cells of which three appear to be from the non-spli ... | 1993 | 8376961 |
| expression of bovine vitamin k-dependent carboxylase activity in baculovirus-infected insect cells. | a vitamin k-dependent carboxylase has recently been purified from bovine liver microsomes and candidate cdna clones have been isolated. definitive identification of the carboxylase remains circumstantial since expression of candidate carboxylase cdnas in mammalian cells is confounded by the presence of endogenous carboxylase activity. to overcome this problem, a recombinant strain of baculovirus (autographa california nuclear polyhedrosis virus, acmnpv) encoding a putative carboxylase (vbcbx/acm ... | 1993 | 8378308 |
| a 37-kilodalton glycoprotein from a baculovirus of orgyia pseudotsugata is localized to cytoplasmic inclusion bodies. | the gene encoding a 37-kda glycoprotein (gp37) of orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) was located and sequenced. gp37 of opmnpv was found to have 62 and 37% amino acid sequence identity with gp37 of autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and with a protein reported to be a component of occlusion bodies from choristoneura biennis entomopoxvirus, respectively. the mrna start site of the opmnpv gp37 gene was mapped within ... | 1993 | 8380088 |
| development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells. | baculovirus multiple gene transfer vectors pacab3 and pacab4 have been developed to facilitate the insertion of three or four foreign genes respectively into the autographa californica nuclear polyhedrosis virus (acnpv) genome by a single co-transfection experiment. the pacab3 vector contains a polyhedrin promoter and two p10 promoters on either side of the polyhedrin promoter but in opposite orientations. the pacab4 vector has an additional polyhedrin promoter in opposite orientation to the fir ... | 1993 | 8385313 |
| the structural and functional organization of the autographa californica nuclear polyhedrosis virus genome. | baculoviruses are used as biological control agents of insect pests in agriculture and forestry. the multiple-nucleocapsid nuclear polyhedrosis virus of autographa californica (acmnpv) is the prototype baculovirus. recently, this virus has become widely used as vector for the high-level expression of foreign genes in insect cells. an updated physical map of restriction sites as well as the location of open reading frames (orfs) and transcripts are presented. most characteristic is the dispersal ... | 1993 | 8389114 |
| nucleotide sequence of the ubiquitin-39k gene region from the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. | the region of the orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (opmnpv) genome containing the ubiquitin and the nuclear matrix-associated protein (39k) genes was sequenced. the first 77 amino acids of the opmnpv ubiquitin open reading frame (orf) showed 84.4% and 79% amino acid sequence identity to autographa californica multinucleocapsid nuclear polyhedrosis virus (acmnpv) and the spodoptera frugiperda insect ubiquitin, respectively. the predicted opmnpv ubiquitin protein c ... | 1993 | 8389803 |
| host range expansion of autographa californica nuclear polyhedrosis virus (npv) following recombination of a 0.6-kilobase-pair dna fragment originating from bombyx mori npv. | we have isolated hybrid baculoviruses of bombyx mori nuclear polyhedrosis virus (bmnpv) and autographa californica npv (acnpv) capable of replicating in both bmn (not susceptible to acnpv) and sf-21 (not susceptible to bmnpv) cells (a. kondo and s. maeda, j. virol. 65:3625-3632, 1991). repeated backcross infection of one of these recombinant isolates with acnpv generated eh-acnpv, a virus with restriction endonuclease patterns of genomic dna nearly identical to those of acnpv but capable of repl ... | 1993 | 8396678 |
| identification and characterization of the p35 gene of bombyx mori nuclear polyhedrosis virus that prevents virus-induced apoptosis. | nucleotide sequence analysis of the bombyx mori nuclear polyhedrosis virus (bmnpv) genome revealed the existence of a gene homologous to the p35 gene of autographa californica npv (acnpv), which has been shown to prevent virus-induced apoptosis. the bmnpv p35 gene showed 96.1% nucleotide and 89.6% predicted amino acid sequence identity to the acnpv p35 gene. a mutant bmnpv (bmp35z) lacking a functional p35 gene induced apoptosis-like cell degradation in infected bmn cells. however, unlike the p3 ... | 1993 | 8416377 |