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primary structure of the s peplomer gene of bovine coronavirus and surface expression in insect cells.the nucleotide sequence of the s peplomer gene of bovine coronavirus (bcv) has been determined. a single open reading frame of 4089 nucleotides encodes a polypeptide of 150k with 20 potential sites for addition of n-linked oligosaccharides. expression of the cloned bcv s gene by a recombinant of autographa californica nuclear polyhedrosis virus resulted in production of a 180k glycosylated polypeptide which was transported to the surface of the cell. comparison of the bcv s gene with the analogo ...19902155283
synthesis of functional bovine leukemia virus (blv) p34tax protein by recombinant baculoviruses.the bovine leukemia virus (blv) p34tax (also called tat, p34, xlor gene product) is a 34-kda polypeptide encoded in the 3'-terminal region of the virus. this protein is responsible for positive transcriptional trans-activation of promoter elements located within the blv long-terminal repeat. we introduced the protein-coding region of blv p34tax into the genome of the baculovirus autographa californica nuclear polyhedrosis virus. after infection of the insect spodoptera frugiperda (sf9) cell line ...19892554576
the antiviral spectrum of (e)-5-(2-bromovinyl)-2'-deoxyuridine.the antiviral activity spectrum of (e)-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) is not restricted to herpes simplex virus type 1 (hsv-1) and varicella-zoster virus (vzv) but also encompasses several other herpesviruses such as suid herpesvirus type 1 (shv-1), bovid herpesvirus type 1 (bhv-1), simian varicella virus (svv), herpesvirus saimiri, herpesvirus platyrrhinae, and the baculovirus trichoplusia ni multiple nuclear polyhedrosis virus. other herpesviruses such as herpes simplex virus type 2, ...19846092320
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 ...19948042283
bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the m2 protein.the coding region of the gene for the nucleocapsid (n) protein of bovine respiratory syncytial virus has been inserted into the genome of autographa californica nuclear polyhedrosis virus (acnpv) using transfer vector pvl 1393. infection of spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of n protein. this protein was indistinguishable from the authentic bovine respiratory syncytial virus (brsv) n protein by sds-gel electrophoresis and immunoprecipitat ...19938438581
expression and one-step purification of bovine interleukin-21 (il-21) in silkworms using a hybrid baculovirus expression system.a hybrid baculovirus, a hybrid of the autographa californica nuclear polyhedrosis virus and the bombyx mori nuclear polyhedrosis virus, was used for the large-scale production of bovine interleukin-21 (il-21) in silkworms. a recombinant hybrid baculovirus containing the full length of the cdna of bovine interleukin-21 was constructed and used to infect silkworm larvae or silkmoth pupae. after the infection of the virus, bovine mature il-21 was produced in the haemolymph or pupal cell lysates. a ...200415604780
high-level expression of recombinant 3ab1 non-structural protein from fmdv in insect larvae.for its potential usefulness in diagnosis, the non-structural protein 3ab1 from foot-and-mouth disease virus was expressed as a soluble protein by using autographa californica nuclear polyhedrosis virus as a vector. the 3ab1 coding sequence was introduced into acnpv genome via pbacpak3ab1 transfer vector to originate ac3ab1 recombinant baculovirus of phenotype occ-. rachiplusia nu larvae were injected with supernatants of sf9 cells infected with ac3ab1 and 5 days post-infection total protein ext ...200415664073
baculovirus treatment fully protects mice against a lethal challenge of fmdv.foot-and-mouth disease virus (fmdv) causes a highly contagious and economically devastating disease that affects cattle, swine, goat and sheep among others. fmdv is able to overcome the initial host innate immune response by inhibiting the induction of antiviral molecules at both the transcriptional and the translational levels. it has been demonstrated that fmdv a/arg/2001 causes the death of adult c57bl/6 mice within 72h. we evaluated the capacity of autographa californica nuclear polyhedrosis ...201020580746
stable protein expression in mammalian cells using baculoviruses.the baculovirus autographa californica multiple nuclear polyhedrosis virus (acmnpv) has been widely used in biotechnology for protein expression in insect cells. baculoviruses use arthropods as their natural hosts and are unable to replicate in mammalian cells. however, acmnpv is able to enter many mammalian cell types and can be used for transgene expression if engineered to contain suitable expression cassettes. in this chapter, we describe the construction and application of a recombinant bac ...201221987248
the effects of bmnpv on biochemical changes in primary cultures of bombyx mori embryonic tissue.the effect of bombyx mori nuclear polyhedrosis virus (bmnpv) on biochemical changes of tc-100 medium containing 10% fetal bovine serum (fbs) in embryonic primary cultures of silkworm was investigated. the primary cultures that reached 60% confluence were infected by 0.5, 1, and 2-ml viral inoculums (diluted with tc-100 medium representing multiplicity of infection (moi) of 0.25, 0.5, and 1). glucose, uric acid, urea, total protein, cholesterol, and alkaline phosphatase were measured in the mediu ...200818288542
expression of bioactive recombinant bovine interferon-gamma using baculovirus.the precise role of bovine interferon-gamma (boifn-gamma) in disease and therapy is still poorly defined. clearly it is involved in defence against parasites, bacteria, viruses and possibly tumor cells. this paper reports the expression of boifn-gamma in a baculovirus system to generate a fully functional recombinant protein. bovine interferon-gamma cdna was cloned from mitogen stimulated peripheral blood mononuclear cell (pbmc) rna utilizing the reverse transcription-polymerase chain reaction ( ...200616608121
establishment and characterization of a continuous cell line from pupal ovaries of japanese oak silkworm antheraea yamamai guerin-meneville.pupal ovaries of the wild oak silkworm antheraea yamamai guerin-meneville were cultured in mgm-448 (modified grace medium-448) medium containing 10% fetal bovine serum. after the primary culture was set up in 1988, a continuous cell line was obtained in 1991, designated as nises-anya-0611 (anya-0611). the population doubling time was 54 hrs. and 19 min. at 96 passages and 88 hrs. and 29 min. at 387 passages. spindle-shaped and spherical cells coexisted in the cell group. the cell line karyotype ...200312892519
production of biologically active recombinant bovine interferon-gamma by two different baculovirus gene expression systems using insect cells and silkworm larvae.the full-length bovine interferon-gamma (bifn-gamma) cdna, including the secretion signal peptide coding region was recloned into baculovirus transfer vectors pacym1 and pbm050. these vectors were co-transfected with autographa californica nuclear polyhedrosis virus (acnpv) or bombyx mori nuclear polyhedrosis virus (bmnpv) dna into spodoptera frugiperda cells (sf21ae) and bombyx mori cells (bmn), respectively. the recombinant viruses, named acbifn-gamma and bmbifn-gamma, were then recovered. rec ...200111145838
the expression and biologic effects of ovine interleukin-4 on t and b cell proliferation.using the reverse-transcriptase polymerase chain reaction (rt-pcr), cdna encoding ovine (ov) interleukin-4 (ovil-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (pbmc). two identical clones generated from separate rt-pcr reactions differed from a published ovil-4 sequence, although they had a high degree of identity with the bovine and human homologs. we show by sequence analysis that the ovil-4 cdna retained the four alpha-helix structure and disulfide bonds identifi ...200010805377
expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (bhv-1) glycoprotein d (gd) sequences.glycoprotein d (gd) of bovine herpesvirus 1 (bhv-1), a homolog of herpes simplex virus gd, represents a major component of the viral envelope and is a dominant immunogen. to study the antigenic properties of the different regions of gd, we have expressed the full-length gd encoding gene and overlapping fragments spanning various regions of the gd open reading frame in a baculovirus (autographa californica nuclear polyhedrosis virus)--insect cell (spodoptera frugiperda, sf-9) system. maximum leve ...19989856100
establishment of a cell line from spodoptera litura (lepidoptera: noctuidae) and replication of s. litura nuclear polyhedrosis virus in vitro.a new cell line, designated ibl-slo1a, was established from pupal ovaries of the tobacco cutworm spodoptera litura. cells were grown in the tnm-fh insect cell culture medium supplemented with 10% fetal bovine serum. the subculture of cells was initiated with 1-3 x 10(5) cells/ml, a split ratio about 1:10. the growth curve of the cells fit the model of exponential growth (y = e-0.6750 + 0.0317x r2 = 0. 98), the population doubling time during logarithmic growth at 28 degrees c was 21.9 hr. the nu ...19979028921
cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf.a cdna encoding a distinct bovine (bo) interferon (ifn) alpha, designated boifn-alpha e, was generated from gut epithelial cells isolated from a rotavirus-infected calf. the boifn-alpha e cdna sequence shared a greater than 90% identity with the other boifn-alpha subtypes. the cdna encoding boifn-alpha e has been expressed in insect cells using the baculovirus autographa californica nuclear polyhedrosis virus (acmnpv) as a vector. insect cells infected with recombinant virus secreted a protein w ...19968640447
construction and insect larval expression of recombinant vesicular stomatitis nucleocapsid protein and its use in competitive elisa.the gene encoding the nucleocapsid (n) protein of indiana 1 serotype vesicular stomatitis virus (vsv-in1) was transferred into the genome of autographa californica nuclear polyhedrosis virus (baculovirus) as a full-length non-fusion construct under the control of the polyhedrin gene promoter. recombinant n protein was obtained from trichoplusia ni insect larvae inoculated 72-96 h previously with the recombinant baculovirus. polyclonal antibody (pab) against vsv-in1 was produced in mice using vsv ...19958530566
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 ...19938378308
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 ...19938347597
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 ...19947530594
expression of human papillomavirus type 6b e2 gene product with dna-binding activity in insect (bombyx mori) cells using a baculovirus expression vector.human papillomavirus type 6b (hpv6b) has been shown to be a major etiologic agent of genital condylomas. the e2 gene, one of the early genes, has been shown to activate the enhancer element in trans in cells transformed with bovine papillomavirus type 1a (bpv1a) but the e2 gene product of any hpv has not been identified. the e2 gene of hpv6b was inserted into the polyhedrin gene of a baculovirus, bombyx mori nuclear polyhedrosis virus (bmnpv), 156 bp downstream from the translational start codon ...19882842232
expression of the e2 open reading frame of papillomaviruses bpv1 and hpv6b in silkworm by a baculovirus vector.to produce and characterize the nature of the e2 protein of papillomaviruses, we have developed a system to express the dna sequence containing an open reading frame (orf) as a fusion protein in cultured insect cells and silkworms. the dna fragments of the e2 orf predicted from the dna sequence of bovine papillomavirus type 1 and human papillomavirus type 6b were linked to the n-terminal part of polyhedrin gene of the baculovirus bombyx mori nuclear polyhedrosis virus (bmnpv) vector. hybrid prot ...19882837019
expression and secretion of the bovine coronavirus hemagglutinin-esterase glycoprotein by insect cells infected with recombinant baculoviruses.a cdna fragment representing the hemagglutinin-esterase (he) gene of bovine coronavirus (bcv) was inserted into the genome of autographa californica nuclear polyhedrosis virus. infection of insect cells with the recombinant virus resulted in the production of a 120-kilodalton disulfide-linked dimeric form of the bcv he polypeptide. deletion of the carboxy-terminal hydrophobic domain from the he polypeptide resulted in secretion of a dimeric form of the truncated he polypeptide. the acetylesteras ...19902181154
alpha 1----3-galactosyltransferase: the use of recombinant enzyme for the synthesis of alpha-galactosylated glycoconjugates.we have reported the isolation and characterization of a bovine cdna clone containing the complete coding sequence for udp-gal:gal beta 1----4glcnac alpha 1----3-galactosyltransferase [joziasse, d. h., shaper, j. h., van den eijnden, d. h., van tunen, a. j. & shaper, n. l. (1989) j. biol. chem. 264, 14290-14297]. insertion of this cdna clone into the genome of autographa californica nuclear polyhedrosis virus (acnpv) and subsequent infection of spodoptera frugiperda (sf9) insect cells with recom ...19902116309
characterization of virus-like particles produced by a recombinant baculovirus containing the gag gene of the bovine immunodeficiency-like virus.the entire gag gene of the bovine immunodeficiency-like virus (biv) was inserted behind the strong polyhedron promoter of autographa californica nuclear polyhedrosis virus (acnpv). the resultant recombinant baculovirus (acnpv-bivgag) was used to infect insect cells in order to overexpress and characterize biv gag gene products. the infection resulted in the high-level expression of a protein similar in size to the predicted biv gag precursor (pr53gag). biv pr53gag was detected in acnpv-bivgag-in ...19901699349
investigation of reduced serum and serum-free media for the cultivation of insect cells (bm5) and the production of baculovirus (bmnpv).an experimental study has been carried out to investigate the effectivenes of several reduced serum and serum-free media for the cultivation of an ovarian cell line, bm5, of the lepidopteran insect bombyx mori. bm5 cell were successfully adapted to grow in a medium containing 5% serum and a serum-free medium (ex-cell 400). on the other hand, this cell line could not be adapted to grow in several other media suggested in the literature, including ipl-41 + 2% fetal bovine serum (fbs), sf-900, and ...199218601067
biological activity of recombinant bovine interferon tau using an autographa californica nuclear polyhedrosis virus expression system.bovine interferon (bifn) tau, which plays a key role in maternal-fetal recognition of pregnancy, was expressed by an autographa californica nuclear polyhedrosis virus expression system. cdna coding bifntau was derived from cultured trophoblast cells. the recombinant (r) bifntau had high antiviral activity (1 x 10 (8) iu/mg) and the molecular weight of rbifntau was estimated to be 23 kda by western blotting analysis. we investigated the biological effect of rbifntau on prostaglandin (pg) f(2alpha ...200314967893
baculovirus transactivator ie1 is functional in mammalian cells.ie1 of autographa californica multicapsid nuclear polyhedrosis virus acts as a transactivator of several viral promoters in insect cells. transient expression assays indicate that ie1 is involved in the activation of the early promoter he65 in both insect tn-368 and mammalian bhk-21 cell lines. ie1 activation of the he65 promoter was compared with ie1 activation of the early 39k promoter. in contrast to the he65 promoter, the 39k promoter was not inducible by ie1 in bhk-21 cells.19979191949
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