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baculovirus replication: inhibition of trichoplusia ni multiple nuclear polyhedrosis virus by [e]-5-(2-bromovinyl)-2'-deoxyuridine.[e]-5-(2-bromovinyl)-2'-deoxyuridine (bvdu) inhibits the replication of the baculovirus trichoplusia ni multiple nucleocapsid nuclear polyhedrosis virus in spodoptera frugiperda cells. virus-specific dna synthesis and late protein synthesis are suppressed by the drug. bvdu is phosphorylated by deoxythymidine (deoxycytidine) kinase present in both uninfected and virus-infected cells, and in its 5'-triphosphate form it inhibits dna polymerase activity in virus-infected cells. the effect of the bvd ...19836343553
safety evaluation of nuclear polyhedrosis virus replication in pigs.to evaluate the hygienic risk involved in using baculoviruses for insect pest control, safety studies are required. pigs were chosen as representative test animals of commercial and agricultural importance. the tests were aimed at detecting virus propagation, immune reactions, and signs of acute infection (changes in body temperature and hematology profile, swelling of lymph nodes). four of five animals inoculated with nuclear polyhedrosis virus showed a slight temperature rise at day 2 postinfe ...19836344789
persistent baculovirus infections: spodoptera frugiperda npv and autographa californica npv in spodoptera frugiperda cells.establishment of a persistent infection of spodoptera frugiperda nuclear polyhedrosis virus (npv) in spodoptera frugiperda (s.f.) cells occurred in three phases: the first phase was characterised by high levels of cell infection and death, the second phase by decreasing cell infection levels leading to the final phase where less than one per cent of the cells were infected during any subculture. the virus persisted at this level of infection provided the cells were maintained by regular subcultu ...19836360080
physical analysis of autographa californica nuclear polyhedrosis virus transcripts for polyhedrin and 10,000-molecular-weight protein.in autographa californica nuclear polyhedrosis virus-infected cells, polyhedrin, the major structural polypeptide of the viral occlusions, and a low-molecular-weight viral protein with a molecular weight of approximately 10,000 (10k) accumulated to high levels late in infection. two polyadenylated rnas 1,200 and 630 bases in size were the most abundant viral transcripts present in the cytoplasm at 48 h postinfection. evidence is presented that these rnas were the transcripts for polyhedrin and 1 ...198316789235
location of homologous dna sequences interspersed at five regions in the baculovirus acmnpv genome.an examination of autographa californica nuclear polyhedrosis virus dna revealed the presence of five interspersed regions, rich in ecori restriction sites, which shared homologous sequences. these homologous regions (hr), designated hr(1) to hr(5), occur at or near the following ecori fragment junctions: hr(1)ecori-b-ecori-i (0.0 map units); hr(2), ecori-a-ecori-j (19.8 map units); hr(3), ecori-c-ecori-g (52.9 map units); hr(4), ecori-q-ecori-l (69.8 map units); and hr(5), ecori-s-ecori-x (88.0 ...198316789237
occluded and budded autographa californica nuclear polyhedrosis virus: immunological relatedness of structural proteins.the immunological relatedness of the structural proteins of the budded and occluded phenotypes of autographa californica nuclear polyhedrosis virus was examined by reciprocal immunoblotting and by in situ peroxidase-antiperoxidase staining of virus-induced cell surface and intracellular antigens with antisera to both phenotypes. the molecular weights (mws) of major structural proteins of both phenotypes that reciprocally cross-reacted were 92,500, 78,000, 62,500, 54,000, and 42,000. a highly imm ...198316789239
protein kinase activity associated with the extracellular and occluded forms of the baculovirus autographa californica nuclear polyhedrosis virus.protein kinase activity is associated with both the extracellular and the occluded forms of autographa californica nuclear polyhedrosis virus, a baculovirus. serine and threonine are the predominant amino acids phosphorylated by the kinase activity associated with both viral forms; no phosphotyrosine was detected. the addition of calcium, camp, or cgmp has no apparent effect on the amount of phosphorylation or the substrates phosphorylated.198316789240
molecular engineering of the autographa californica nuclear polyhedrosis virus genome: deletion mutations within the polyhedrin gene.we describe a method to introduce site-specific mutations into the genome of autographa californica nuclear polyhedrosis virus. specifically, the a. californica nuclear polyhedrosis virus gene for polyhedrin, the major protein that forms viral occlusions in infected cells, was mutagenized by introducing deletions into the cloned dna fragment containing the gene. the mutagenized polyhedrin gene was transferred to the intact viral dna by mixing fragment and viral dnas, cotransfecting spodoptera fr ...198316789242
viral transcription during autographa californica nuclear polyhedrosis virus infection: a novel rna polymerase induced in infected spodoptera frugiperda cells.autographa californica nuclear polyhedrosis virus-specific rna synthesis in isolated nuclei of spodoptera frugiperda cells in culture was monitored at different times postinfection. up to 8 h postinfection viral rna synthesis remained sensitive to 5 mug of alpha-amanitin per ml. during the course of infection this sensitivity decreased, and at 24 h postinfection rna synthesis was completely resistant to alpha-amanitin. deae-sephadex profiles of rna polymerase isolated at 24 h postinfection showe ...198316789246
characterization of two abundant mrnas of autographa californica nuclear polyhedrosis virus present late in infection.cytoplasmic poly(a)(+) rna isolated from spodoptera frugiperda cells late after infection with autographa californica nuclear polyhedrosis virus (30-40 hr pi) was fractionated according to size on denaturing methyl mercury gels. two major rna species (1.4 kb and 0.75 kb) and several minor rna species were detected by ethidium bromide staining. the predominant rna species of about 1.4 kb was considered to be polyhedrin mrna because (1) in vitro translation of the rna, which was eluted from methyl ...198318638868
a temperature-sensitive mutant of the baculovirus autographa californica nuclear polyhedrosis virus defective in an early function required for further gene expression.fifteen temperature-sensitive (ts) mutants of the baculovirus autographa californica nuclear polyhedrosis virus (acnpv) have been analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) of proteins synthesized in infected cells. one of the mutants, tsb821, was found to be defective in a very early function. seven virus-induced proteins were synthesized by 2 hr postinfection. in marked contrast to wild-type virus and the other 14 ts mutants, the synthesis of further virus ...198318638939
mapping early transcription products of autographa californica nuclear polyhedrosis virus.the regions of the acmnpv genome represented as cytoplasmic transcripts early after infection of spodoptera frugiperda cells prior to and encompassing the initiation of dna replication were mapped. in vivo(32)p pulse-labeled cytoplasmic rna from infected cells at various times postinfection was used to probe southern blots of cloned ecori acmnpv dna fragments. at the earliest time point studied (0.5 - 2.5 hr p.i.) transcripts represented a large proportion of the genome although specific regions ...198318638940
a semipermissive nuclear polyhedrosis virus infection: characterization of infection kinetics and morphogenesis.an in vitro host-range model system, designated system ii, was developed for the multicapsid nuclear polyhedrosis virus spodoptera frugiperda (sf-mnpv). this system consisted of sf-mnpv-infected s. frugiperda cells (permissive system) and sf-mnpv-infected trichoplusia ni cells (semipermissive system). infection kinetic studies revealed that while progeny virus was produced in the semipermissive system, the infectious virus yield was at least 700-fold lower than that in positive controls. electro ...198318639138
a study of the glycoproteins of autographa californica nuclear polyhedrosis virus (acnpv).pulse labeling with tritiated mannose was used to follow the time course of autographa californica nuclear polyhedrosis virus (acnpv) glycoprotein synthesis in spodoptera frugiperda iplb-21 cells. nine viral-induced intracellular glycoproteins were first detected from as early as 2 hr postinoculation (67k, early phase) to as late as 14 hr (36k and 19k glycoproteins, intermediate phase). glycosylation of these proteins was observed to continue to the end of the experiment (28 hr postinoculation). ...198318639173
nucleotide sequence of the polyhedrin gene of autographa californica nuclear polyhedrosis virus.the nucleotide sequence of a 1143-bp region of autographa californica mnpv (acmnpv) dna containing the entire coding region of polyhedrin was determined. the coding region consisted of 732 bp without intervening sequences. the 5' leader sequence was approximately 58 nucleotides in length. a putative "tata box" and "caat box" were located 20 and 52 bp upstream from the transcriptional start site, respectively. two tandem repeats of 8 bp were found 70 and 84 bp upstream from the transcriptional st ...198318639177
dynamics of baculovirus growth and dispersal in mamestra brassicae l. (lepidopteranoctuidae) larval populations introduced into small cabbage plots.the nuclear polyhedrosis virus of mamestra brassicae has been studied in larval populations of the moth introduced into small plots of cabbages. primary dispersal of virus from single foci of infected larvae resulted from enhanced movement of the larvae, which colonized new plants logarithmically. virus growth within the host population was quantified, and infection of young larvae in the following generation was related directly to the concentration of virus produced during the primary phase. s ...198316346197
in vitro survey of autographa californica nuclear polyhedrosis virus interaction with nontarget vertebrate host cells.thirty-five nontarget host cell lines, 23 of human and 12 of nonhuman vertebrate origin, were exposed to autographa californica nuclear polyhedrosis virus preparations derived from four different sources: polyhedra, hemolymph, cell culture medium, and cultured cells. the virus and cells were incubated together at two different temperatures, 28 or 37 degrees c, for four different lengths of time, 16, 40, 64, or 168 h, and the cells were assayed for the presence of virus by a peroxidase-antiperoxi ...198316346229
characterization of gypsy moth (lymantria dispar) nuclear polyhedrosis virus.characterization of the proteins and nucleic acid of the gypsy moth nuclear polyhedrosis virus isolated in ithaca, n.y. (ldnpv-it) is presented. a total of 29 viral structural proteins were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis when the virus was isolated in the absence of alkaline protease activity. fourteen surface envelope viral proteins were identified by lactoperoxidase iodination. eleven proteins were associated with nucleocapsids prepared by nonidet p-40 ...198316346355
two naturally occurring nuclear polyhedrosis virus variants of neodiprion sertifer geoffr. (hymenoptera; diprionidae).two isolates of neodiprion sertifer (geoffr.) nuclear polyhedrosis virus from britain and north america were compared using three biochemical techniques. alkaline protease assays of polyhedra revealed the presence of endogenous enzyme activity in the british isolate but not in the north american isolate. sodium dodecyl sulfate polyacrylamide gel electrophoresis of virus particle structural polypeptides revealed only minor differences, with the exception that the north american virus was contamin ...198216345930
generalized immunoassay for autographa californica nuclear polyhedrosis virus infectivity in vitro.a quantitative in vitro immunoassay for the infectivity of autographa californica nuclear polyhedrosis virus was developed and performed in six different lepidopteran cell lines. the assay was not dependent upon cytopathic effect or polyhedron production, but rather upon viral antigen production and its recognition in a peroxidase-antiperoxidase staining procedure. the importance of using such an assay for accurately assessing infectivity in cell lines which produce polyhedra inefficiently was d ...198216346059
orientation of the genome of autographa californica nuclear polyhedrosis virus: a proposal.the nuclear polyhedrosis virus of the alfalfa looper autographa californica contains a double-stranded, circular dna genome. fourteen scientists agreed to accept an orientation of this circular genome with respect to a physical map of the restriction endonuclease cleavage sites.198216789218
molecular cloning and physical mapping of restriction endonuclease fragments of autographa californica nuclear polyhedrosis virus dna.a restriction fragment library containing autographa californica nuclear polyhedrosis virus (acnpv) dna was constructed by using the pbr322 plasmid as a vector. the library, which is representative of more than 95% of the viral genome, consists of 2 of the 7 bamhi fragments, 12 of the 24 hindiii fragments, and 23 of the 24 ecori fragments. the cloned fragments were characterized and used to generate physical maps of the genome by hybridizing nick-translated recombinant plasmid to southern blots ...198216789219
capping of viral rna in cultured spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus.viral rna from fall armyworm (spodoptera frugiperda) cells infected with autographa californica nuclear polyhedrosis virus contains cap structures. most of the cap labeled in vivo with [(3)h]methionine or (32)p(i) cochromatographed on deae-cellulose with the -5 charge marker; a minor component appeared at -4 net charge. the former is probably a cap 1 structure (m(7)gpppx(m) (p)yp), and the latter is probably a cap 0 (m(7)gpppxp). on the basis of relative labeling of the two caps with [(3)h]adeno ...198216789227
mapping the mutation site of an autographa californica nuclear polyhedrosis virus polyhedron morphology mutant.a polyhedron morphology mutant of autographa californica nuclear polyhedrosis virus, designated m5, was compared with wild-type virus by genotypic analysis with ecori, bamhi, hindiii, ssti, and smai restriction endonucleases. m5 dna revealed several alterations relative to the wild-type pattern: (i) ecori fragment i was 400 base pairs larger; (ii) bamhi fragment f was missing; (iii) hindiii fragment f was 400 base pairs larger; (iv) an extra restriction fragment was obtained with both hindiii an ...198216789229
in vitro translation of autographa californica nuclear polyhedrosis virus early and late mrnas.a preliminary translational map of the autographa californica genome was constructed. eighteen viral dna restriction fragments were either purified from agarose gels or obtained from pbr322 recombinant dna plasmids to locate specific gene products. the dnas were immobilized on nitrocellulose filters and used to select viral mrnas isolated from rna obtained from the cytoplasm of infected spodoptera frugiperda cells at 21 h postinfection. the fragment-specific mrnas were translated in vitro in the ...198216789231
analysis of the spodoptera frugiperda nuclear polyhedrosis virus genome by restriction endonucleases and electron microscopy.restriction endonuclease analysis was used to differentiate between four strains of spodoptera frugiperda nuclear polyhedrosis virus from different geographical areas. in addition, partial denaturation was performed, and a partial denaturation map was constructed for the ohio strain of this virus.198216789233
rifampin inhibition of the occluded virus form of a nuclear polyhedrosis virus.rifampin at concentrations toxic to noninfected cells but not to infected cells is a selective inhibitor of occluded virus of the group a baculoviridae (nuclear polyhedrosis virus). however, the titer of nonoccluded virus is not affected. rifampin blocks occlusion until late in the replication cycle (14 to 16 h), and its effects are reversible. modes of action of polyhedral inclusion body production are unknown.19826291455
properties of the nuclear polyhedrosis virus of the great wax moth: oligomeric circular dna and the characteristics of the genome. 198218638732
identification, cloning, and r-loop mapping of the polyhedrin gene from the multicapsid nuclear polyhedrosis virus of orgyia pseudotsugata.polyadenylated rna was isolated from orgyia pseudotsugata larvae 8-10 days postinfection with the multicapsid nuclear polyhedrosis virus. this rna was centrifuged through a sucrose gradient and fractions enriched for polyhedrin mrna were identified by in vitro translation. complementary dna made to this rna hybridized predominantly to a 5-kb fragment of xhoi-digested viral dna. this fragment was cloned into the plasmid pacyc177 and mapped with restriction endonucleases. a sali subclone with a 2. ...198218638748
characterization of monoclonal antibodies to the autographa californica nuclear polyhedrosis virus.mouse hybridomas which produce monoclonal antibodies to the autographa californica nuclear polyhedrosis virus (acnpv) have been prepared and the protein to which each is directed has been determined. the antibody produced by clones 3f3 and 5e9 specifically bound polyhedron protein of mw 32,000 when used in immunoadsorption chromatography when used in elisa they could not distinguish between insect- and cell culture-derived polyhedron protein and also reacted with a wide range of baculoviruses. t ...198218638810
dna homology among subgroup a, b, and c baculoviruses.the dna homology among 18 baculoviruses from subgroups a, b, and c was compared using restriction endonuclease analysis, dna hybridization, and southern blot hybridization. restriction enzyme patterns for each of the viral dnas compared was unique. viral dna homology was evaluated by hybridizing 32p-labeled viral dnas to unlabeled dnas immobilized on nitrocellulose filters. hybridization conditions were varied by using a range of formamide concentrations during annealing. the apparent dna homolo ...198218638848
the isolation and characterization of the mp and fp plaque variants of galleria mellonella nuclear polyhedrosis virus.two plaque morphologies were detected in assays of galleria mellonella nuclear polyhedrosis virus preparations which had been serially passaged in the tn-368 cell line. the fp (few polyhedra) plaques were larger (0.7-mm average diameter) and the infected cells contained less than 10 polyhedra each. the mp (many polyhedra) plaques were smaller (0.5-mm average diameter), and the infected cells contained greater than 10 each. only the fp variant could be purified by plaque picking and remained pure ...198218635122
dna restriction-pattern differences from geographic isolates of spodoptera littoralis nuclear polyhedrosis virus.two nuclear polyhedrosis viruses were isolated from spodoptera littoralis larvae collected from several areas in israel. the two viruses were characterized and compared by restriction endonuclease cleavage of their dna and southern blot hybridization. the two viruses have no detectable sequence homology. a variant, homologous to the more common virus isolate, was also found.198218635146
alteration of autographa californica nuclear polyhedrosis virus dna upon serial passage in cell culture.plaque-purified autographa californica nuclear polyhedrosis virus, acmnpv e2 isolate, was propagated for 30 generations by serial passage in tn-368 cells. after passage, the dnas from 7 of 20 replaque-purified isolates were found to have additional ecori restriction enzyme fragments. these 7 isolates were of three types. the additional ecori fragments of each isolate hybridized to existing viral dna. restriction enzyme mapping with saci and smai show that isolates of each type contain an inserti ...198218635147
inhibition of autographa californica nuclear polyhedrosis virus replication in high-density trichoplusia ni cell cultures.replication of the autographa californica nuclear polyhedrosis virus was studied in trichoplusia ni (tn-368) tissue culture cells under cell density conditions which approximated a confluent monolayer (5.7 x 10(5) cells/cm(2) growth surface). these high-density cultures were not refractory to infection but the synthesis of both the occluded and nonoccluded forms of the virus were inhibited greater than 98% when compared to inoculation of low cell density cultures (1.4 x 10(5) cells/cm(2)). the i ...198218635148
the translational map of the autographa californica nuclear polyhedrosis virus (acnpv) genome.we have mapped early and late viral gene products expressed in autographa californica nuclear polyhedrosis virus ( acnpv )-infected spodoptera frugiperda cells by cell-free translation of virus-specific rna which was selected by hybridization to cloned restriction endonuclease fragments of acnpv dna. proteins synthesized in vitro were labeled with [35s]methionine and analyzed by sds-polyacrylamide gel electrophoresis followed by fluorography. at least four early acnpv -specific polypeptides were ...19826327284
molecular cloning of dna complementary to mrna of the baculovirus autographa californica nuclear polyhedrosis virus: location and gene products of rna transcripts found late in infection.dnas complementary to late autographa californica nuclear polyhedrosis virus (acnpv) mrna were synthesized by reverse transcription and cloned in escherichia coli by using pbr322 as a vector. eleven different cdnas were distinguished in our screening of 45 acnpv-homologous clones. location of the regions of cdna homology with respect to the acnpv physical map showed that the 11 cdnas were dispersed throughout the genome. the most abundant cdna insertion, representing approximately one-third of t ...19826184487
bromodeoxyuridine-induced mutants of autographa californica nuclear polyhedrosis virus defective in occlusion body formation.twelve temperature-sensitive (ts) and two morphology mutants of autographa californica nuclear polyhedrosis virus were generated using the mutagen 5-bromo-2'-deoxyuridine (brdurd). the ts mutants grew normally at 25 degrees c but exhibited abnormal occlusion body formation when grown at 33 degrees c whereas the morphology mutants produced an abnormal cytopathic effect at 25 degrees c and 33 degrees c. none of the mutants was severely restricted for production of non-occluded virus; thus, with th ...19826754865
the use of a protein a conjugate in an indirect enzyme-linked immunosorbent assay (elisa) of four closely related baculoviruses from spodoptera species.the virus particles from four closely related insect nuclear polyhedrosis virus (npvs) isolated from their homologous spodoptera spp. hosts were examined using an indirect enzyme-linked immunosorbent assay (elisa), employing both swine anti-rabbit and protein a conjugate. results from both systems indicated that s. littoralis npv is serologically distinct from the other three viruses, s. frugiperda npv, s. exempta npv and s. exigua npv, which are closely related. the relatedness of these viruses ...19826754866
transcription of the autographa californica nuclear polyhedrosis virus genome: location of late cytoplasmic mrna. 19826755883
effect of aluminum chloride and zinc sulfate on autographa california nuclear polyhedrosis virus (acnpv) replication in cell culture.when ipl-sf-21ae iii continuous insect cell line was grown and maintained in ipl-41 insect cell culture medium supplemented with 16 microm of alcl3 or 0.24 microm of znso4 . 7h2o, or both metallic salts, and then infected with autographa california nuclear polyhedrosis virus, virus replication was increased significantly. the yield of polyhedral inclusion bodies (pib) was enhanced up to 121%. synthesis of cell-free nonoccluded virus was increased to 365% when infectivity was assayed by the plaqu ...19826759370
[transfection conditions for baculovirus dna in an insect cell line].the following conditions were found to be necessary to achieve infection of a spodoptera frugiperda cell line with purified dna of nuclear polyhedrosis virus of spodoptera exempta (npv): (1) cell monolayers shall be prepared one day before the experiment; the number of cells added to 35-mm wells of plastic clusters (falcon) shall be 1.0 x10(6)-1.1x10(6), and the cells shall be in the stage of logarythmic growth; dna shall be diluted in hebs-buffer (ph 6.95-7.05); the volume of the mixtures added ...19827090350
plaque assay of heliothis zea baculovirus employing a mixed agarose overlay.the nuclear polyhedrosis virus of heliothis zea has been titrated in heliothis zea cells by the plaque method, using 1 percent mixed agarose containing a mixture of seakem and ultra pure agarose. visible plaques, formed 8 days postinfection, ranged in diameter from 0.5 to 2 mm. dose-response experiments indicated that a single particle initiated the formation of a plaque. the titration of heliothis zea baculovirus by the newly described plaque method provides an accurate technique for the determ ...19817213016
effects of red-headed in sawfly, neodiprion lecontei, nuclear polyhedrosis virus on rainbow trout, salmo gairdneri, and daphnia pulex.the effects of a nuclear polyhedrosis virus (npv) of the red-headed pine sawfly, neodiprion lecontei, on rainbow trout, salmo gairdneri, were investigated. the fish were exposed to this virus by intubation and topical application and no ill-effects were observed. similarly, no ill-effects were detected in daphnia pulex when the same npv was added to their culture medium. the materials were lyophilized, npv-infected sawfly larvae (normally used for insect control), lyophilized, uninfected larvae ...19817026655
improved replication of autographa californica nuclear polyhedrosis virus in roller bottles: characterization of the progeny virus.a reproducible growth curve was established for the propagation of autographa californica (speyer) nuclear polyhedrosis virus (acmnpv) in a continuous insect line from spodoptera frugiperda (j. e. smith) during large-volume production. a newly developed method for quantitation of polyhedra inclusion bodies (pib) by electron microscopy (em) during the growth cycle was compared to hemocytometer counts. the virus particles (vp) ratio to pfu and vp per pib were established by em methods. optimal yie ...19817028679
baculovirus bioassay not dependent upon polyhedra production.an immunoperoxidase-based quantitative assay has been developed for infectious units of autographa californica nuclear polyhedrosis virus infecting tn-368 cells. major benefits of this assay over existing quantitative assays for baculoviruses are that it is not dependent upon polyhedra production, it is clear-cut and easy to score, and results are obtained within 24 h.19817028916
growth of nuclear polyhedrosis virus in larvae of the cabbage moth, mamestra brassicae l.growth of nuclear polyhedrosis virus (npv) in 5 larval instars of cabbage moth, mamestra brassicae, has been quantified using 2 methods. numbers of polyhedra were estimated by light microscope counts while concentrations of virus protein antigen were estimated using elisa. virus growth was rapid initially but slowed during its later stages, although elisa protein concentrations decreased once a peak had been reached. there was a linear correlation between polyhedral counts and virus protein duri ...19817034687
baculovirus induction of a dna polymerase.the baculovirus, autographa california nuclear polyhedrosis virus, induced a new aphidicolin-sensitive, alpha-like, dna polymerase upon infection of the lepidopteran noctuid, trichoplusia ni. the new virus-induced dna polymerase could be separated from the host alpha-like polymerase by phosphocellulose chromatography. the two polymerases differed in their sensitivities to heat inactivation, high salt concentrations, and 0.1 m phosphate buffer.19816793735
biological and biochemical investigations on five european isolates of mamestra brassica nuclear polyhedrosis virus.five multiply enveloped european isolates of mamestra brassicae nuclear polyhedrosis virus (oxford, german, french, dutch and danish) were found to be very closely related serologically using the enzyme linked immunosorbent assay (elisa) double antibody sandwich method and immunodiffusion. by sds-polyacrylamide gel electrophoresis of viral proteins and restriction endonuclease analysis of dna using seven enzymes there appeared to be two variants as the oxford and german isolates were distinct fr ...19816271101
replication of heliothis zea baculovirus in an insect cell line.the baculovirus, heliothis zea nuclear polyhedrosis virus (hzsnpv), was grown in the h. zea cell line, iplb-1075. light microscopy observations showed that 95% of inoculated cells had cpe by 6 days postinoculation (p.i.). growth curves of intracellular and extracellular virus showed that maximum titers (tcid50 units) were reached 3-4 days p.i. studies on hzsnpv morphogenesis revealed that ultrastructural events were typical of baculovirus replication in vitro. ecori restriction enzyme analyses o ...19816276328
the cloned ecori fragments of autographa californica nuclear polyhedrosis virus dna. 19816282701
virus envelope acquisition of nuclear polyhedrosis virus in vitro.there have been several reports concerned with the replication and morphogenesis of insect baculoviruses during the past decade [1--7]. while there is general agreement as to the assembly of the virus on the basis of electron microscopic studies, there are still questions regarding the details of the replicative mechanisms, such as the acquisition of the virus envelope. three possible ways have been proposed to describe envelope formation of nuclear polyhedrosis virus: (i) acquisition of budding ...19817018096
in vivo pathway of autographa californica baculovirus invasion and infection.the pathway of autographa californica nuclear polyhedrosis virus (acnpv) infection in cabbage looper, trichoplusia ni, larval midgut cells was studied by ultrastructural and virus titration methods. enveloped virions interacted with microvilli of columnar cells resulting in apparent fusion of the viral envelope and microvillus membrane. after entry into the cell cytoplasm, the intact nucleocapsids appeared to enter the nucleus through nuclear pores, and uncoating of the viral genome took place i ...198118635031
autographa californica nuclear polyhedrosis virus phosphoproteins and synthesis of intracellular proteins after virus infection.polypeptides and phosphoproteins isolated from nuclear and cytoplasmic fractions of tn-368-10 cells infected with autographa californica nuclear polyhedrosis virus (acmnpv) were analyzed by polyacrylamide gel electrophoresis. likewise, acmnpv extracellular virus and alkali-released virus were compared. acmnpv extracellular virus possessed at least nine phosphoproteins while the alkali-released virus had about 14 phosphoproteins. the major structural polypeptide of acmnpv, polyhedrin, was phospho ...198118635034
investigation of genetic heterogeneity in wild isolates of spodoptera frugiperda nuclear polyhedrosis virus by restriction endonuclease analysis of plaque-purified variants.spodoptera frugiperda nuclear polyhedrosis virus (sfmnpv was obtained from four different sources and each isolate was cultured in iplb-sf21 cells. viral dna from each isolate was analyzed with ecor1, xho1, and bamh1 restriction endonucleases. all four isolates could be distinguished on the basis of minor differences in ecor1 patterns. submolar dna restriction fragments were seen in all isolates indicating that each isolate consisted of a mixture of different genotypes. to investigate this, two ...198118635067
cell culture studies with the imc-hz-1 nonoccluded virus.studies were conducted on an adventitious agent (hz-1v) isolated from the imc-hz-1 cell line. it appeared identical to the virus first obtained by granados et al (r. r. granados, t. nguyen, and b. cato, 1978, intervirology, 10, 309-317) from a persistent infection of this cell line. restriction endonuclease digestion of hz-1v dna indicated the agent was different from the s nuclear polyhedrosis virus (hzsnpv) of the host species, heliothis zea, from which the imc-hz-1 cell line was derived by hi ...198118635105
alpha-amanitin-resistant viral rna synthesis in nuclei isolated from nuclear polyhedrosis virus-infected heliothis zea larvae and spodoptera frugiperda cells.[(3)h]rna was synthesized in nuclei isolated at various times postinfection from the fat bodies of heliothis zea larvae infected with h. zea nuclear polyhedrosis virus and from cultured spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus. to detect virus-specific rna synthesis, the [(3)h]rna was hybridized to denatured viral dna immobilized on nitrocellulose filters. nuclear polyhedrosis virus-specific rna synthesis in the infected nuclei isolated from h. ...198116789208
spodoptera frugiperda nuclear polyhedrosis virus genome: physical maps for restriction endonucleases bamhi and hindiii.the physical map for the genome of spodoptera frugiperda nuclear polyhedrosis virus was constructed for restriction endonucleases bamhi and hindiii. the ordering of the restriction fragments was accomplished by cross-blot hybridization of bamhi, hindiii, and ecori fragments. the alignment of the hindiii fragments within the bamhi map was achieved by double digestion with the two restriction endonucleases followed by cross-blot hybridization. the results showed that the viral genome consisted of ...198116789209
construction of a genetic map of the baculovirus autographa californica nuclear polyhedrosis virus by marker rescue of temperature-sensitive mutants.mutations of seven temperature-sensitive mutants of the baculovirus autographa californica nuclear polyhedrosis virus (npv) were mapped with respect to the physical restriction map of the a. californica npv dna by marker rescue. dnas from two distantly related npvs of the multiply embedded type and two npvs of the singly embedded type were unable to rescue two a. californica npv mutants.198116789213
hybridization selection and in vitro translation of autographa californica nuclear polyhedrosis virus mrna.we isolated polyadenylated rna from the cytoplasm of cells infected with autographa californica nuclear polyhedrosis virus late after infection (21 h postinfection). at that time intracellular protein synthesis was directed almost exclusively toward infected cell-specific proteins. the polyadenylic acid-containing rna sequences in the cytoplasm at 21 h postinfection were radiolabeled in vitro and hybridized to a. californica nuclear polyhedrosis virus dna restriction fragments. the polyadenylic ...198116789215
physical factors that affect in vitro autographa californica nuclear polyhedrosis virus infection.of the physical parameters tested for in vitro baculovirus infection, multiplicity of infection was most important in governing percent cell infection. most plaques formed within the first 5 min of incubation. efficiency of infection, however, was low, and the virus titer did not diminish during prolonged incubation. efficiency of infection improved markedly when cells or virus were preincubated with selected polyanions and polycations. precise regulation of the ph, osmotic pressure, and ionic c ...198116345768
characterization of a nuclear polyhedrosis virus isolated from diseased gonometa podocarpi (lepidoptera:lasiocampidae).gonometa podocarpi is an important pest of several species of pine in east africa, and large numbers of trees in plantations in kenya were partially or completely defoliated by the larval stage of this insect. after the infestation in the mt. elgon region, large numbers of dead and moribund larvae were found on the ground. examination of extracts of these larvae demonstrated the presence of an occluded virus. electron microscopy of purified sectioned polyhedra demonstrated the presence of virus ...198116345830
restriction map of rachiplusia ou and rachiplusia ou-autographa californica baculovirus recombinants.the restriction sites of rachiplusia ou nuclear polyhedrosis virus (romnpv) dna were mapped for the endonucleases smai, kpni, bamhi, saci, xhoi, and ecori. of the 60 dna restriction sites of romnpv, 35 mapped in similar positions as compared to the restriction sites of autographa californica nuclear polyhedrosis virus (acmnpv) dna. two plaque-purified viruses, obtained from randomly picked plaques of a wild-type isolate of romnpv, were recombinants of romnpv and acmnpv. the recombinants were sho ...198016789188
comparison of the structure of c- and n-polyhedrins from two occluded viruses pathogenic for orgyia pseudotsugata.c-and n-polyhedrins from a cytoplasmic polyhedrosis virus (a double-stranded rna virus) and a nuclear polyhedrosis virus (a dna virus), respectively, of orgyia pseudotsugata were compared. although both polyhedrins appear to stabilize their respective virions and have similar molecular weights, they differed in amino acid composition, tryptic peptide elution profiles from a cation-exchange resin, and n-terminal amino acid sequence and showed no antigenic relatedness. this suggests that these two ...198016789195
physical maps of autographa californica and rachiplusia ou nuclear polyhedrosis virus recombinants.tn-368 cells were infected simultaneously with the closely related autographa california (acmnpv) and rachiplusia ou (romnpv) nuclear polyhedrosis viruses. progeny viral isolates were plaque purified, and their dnas were analyzed with restriction endonucleases. of 100 randomly cloned plaques, 7 were acmnpv and romnpv recombinants, 5 were romnpv, and 88 were acmnpv. the recombinants contained dna sequences derived from both parental genomes. by comparing the restriction cleavage patterns of paren ...198016789198
on the origin of the polyhedral protein of autographa californica nuclear polyhedrosis virus. isolation, characterization, and translation of viral messenger rna.virus-specific messenger rna was isolated 24 hr postinfection from spodoptera frugiperda cells infected with autographa californica nuclear polyhedrosis virus (acnpv) by means of hybridization with viral dna that was coupled to cellulose. viral mrna was analyzed on isokinetic sucrose gradients and on polyacrylamide gels containing 98% formamide. viral mrna was also translated in an in vitro protein-synthesizing system derived from wheat germ. one product comigrated with purified acnpv polyhedral ...198018631636
autographa californica nuclear polyhedrosis virus-induced proteins in tissue culture.the multiplication of autographa californica nuclear polyhedrosis virus was studied in trichoplusia ni and spodoptera frugiperda insect cell lines. using pulse labeling, 18 viral-induced proteins were identified by polyacrylamide gel electrophoresis. based on infectivity curves and protein synthesis studies, replication could be divided into four phases. during the early phase of infection, four viral-induced proteins with molecular weights of 45, 35, 34, and 31 thousand daltons could be detecte ...198018631644
structural analysis of the matrix protein from the nuclear polyhedrosis virus of heliothis zea.polyhedrin from the nuclear polyhedrosis virus of heliothis zea was analyzed. alkali-solubilized polyhedrin consists of a 12 s aggregate of 27,000 mw subunits. results from chemical crosslinking experiments suggest that 12 subunits are present in the 12 s aggregate. in isoelectric focusing gels, the aggregate migrates as a single entity with an isoelectric point of 5.9. under denaturing conditions, four charge isomers of the subunits are revealed. the presence of alkaline protease activity in th ...198018631649
protease degradation of autographa californica nuclear polyhedrosis virus proteins.the autographa californica nuclear polyhedrosis virus (ac-npv) occlusion bodies produced in vivo contained a protease which degraded occlusion body matrix protein and polypeptides of enveloped nucleocapsids during alkaline dissolution. the protease activity was required for complete dissociation of occlusion body matrix protein from the viral membrane during alkali liberation of the virus from occlusion bodies. in addition protease activity was required for the subsequent removal of the viral me ...198018631654
protein synthesis in cells infected by autographa californica nuclear polyhedrosis virus (ac-npv): the effect of cytosine arabinoside.twenty-two virion polypeptides (vp) were detected reproducibly when the occluded form (ov) of [35s]methionine-labeled ac-npv, purified from polyhedral inclusion bodies (pib), was analyzed by polyacrylamide gel electrophoresis and autoradiography. ten of the vps and the polyhedral protein (pp) were phosphorylated and 6 vps were glycoproteins. purified, nonoccluded virus (nov) revealed 25 polypeptides. during a single cycle of virus replication, the synthesis of 33 infected cell polypeptides (1cp) ...198018631655
isolation and replication of an occlusion body-deficient mutant of the autographa californica nuclear polyhedrosis virus.a spontaneous mutant of the autographa californica nuclear polyhedrosis virus which was deficient in viral occlusion body formation was studied in trichoplusia ni tissue culture cells. virus multiplication could be divided into four phases based on growth curves and the sequential appearance of 25 viral-induced proteins identified by pulse labeling and polyacrylamide gel electrophoresis. during the latent phase, five viral-induced proteins with molecular weights of 68, 47, 35, and 30 thousand (k ...198018631676
physicochemical properties and location of capsule components, in particular the synergistic factor, in the occlusion body of a granulosis virus of the armyworm, pseudaletia unipuncta.a synergistic strain of granulosis virus (gvh) enhances the infection of larvae of the armyworm, pseudaletia unipuncta, with a nuclear polyhedrosis virus (npv). the matrix of the capsule (occlusion body) of gvh contains two major components, the structural protein and the synergistic factor. the structure of the capsule matrix of gvh was analyzed with a primary amine-coupling reagent, 2,4,6,-trinitrobenzene 1-sulfonic acid (tnbs). when undissolved capsules are incubated with tnbs, only the struc ...198018631836
infectious dna from autographa californica nuclear polyhedrosis virus.cells from the lepidopteran spodoptera frugiperda can be successfully transfected in culture with the dna from autographa californica nuclear polyhedrosis virus (acnpv). the calcium chloride precipitation procedure has been used in conjunction with dimethyl sulfoxide treatment of the transfected cells. the highest specific infectivity observed was 6.1 x 10(4) pfu/mug of acnpv dna. as judged from a comparison of the restriction patterns of viral dna preparations, the virus produced in transfectio ...198016945844
[pathohistological and ultrastructural studies of the nuclear polyhedrosis virus in the silkworm (bombyx mori l.)].histopathologic studies were carried out on fifth instar silkworm moths of the super 1 x super 2 hybrid infected orally with baculovirus bombycis along with ultrastructural investigations of the pathogen itself. it was found that the earliest changes in the moths set in at the 60th hour post inoculation. they consisted in the aggregation of chromatin, and the formation of polyhedral bodies in the fatty body, hypodermis, and peritracheal matrix; these appeared in the glandular tissue too, but not ...19807020236
chemically-defined media for production of insect cells and viruses in vitro.two chemically-defined media are described. they support the growth of a) an established cell line of spodoptera frugiperda cells and b) two established mosquito cell lines from aedes aegypti and anopheles gambiae. the replication of autographa californica nuclear polyhedrosis virus (acnpv) in s. frugiperda cells grown in a defined medium is reported.19806102531
virulence of cloned variants of autographa californica nuclear polyhedrosis virus.the relative virulence of five different genotypic variants of autographa californica nuclear polyhedrosis virus was tested by determining the 50% lethal dose of occluded virus for larvae of trichoplusia ni. the 50% lethal dose values of uncloned virus and the five cloned genotypic variants ranged between 10 and 21 polyhedra per larva, and no statistically significant differences were observed. cloning has therefore neither enhanced nor decreased the virulence of this potential microbial pestici ...19806990868
the infectivity of polyhedra of nuclear polyhedrosis virus (n.p.v.) after passage through gut of an insectpredator.polyhedral inclusion bodies (p.i.b) of a nuclear polyhedrosis virus of heliothis punctigera (lep.) were found in excreta of nabis tasmanicus (het.), a predator of h. punctigera. an immunofluorescent counting method was used to estimate numbers of p.i.b. bioassays demonstrated that no loss of infectivity occurred during passage through the gut.19806995145
[degradation of silkworm nuclear polyhedrosis virus]. 19806995813
biological studies of a baculovirus in a mammalian cell line.in two of five experiments, replication (tcid50) was observed following inoculation of chinese hamster cells with autographa californica nuclear polyhedrosis virus (npv). radiolabeling experiments employing tritiated thymidine consistently resulted in a labeled entity which banded in sucrose gradients at a density of 1.24-1.25 g/ml, similar to that observed for a. californica npv in the permissive trichoplusia ni cell line. the labeled entity is believed to be a. californica npv, based on infect ...19806998919
[quantitative determination of the infectivity of nuclear polyhedrosis virus dna on honeycomb moth (galleria mellonella) larvae].the possibility of using honeycomb moth larvae for titration of nuclear polyhedrosis virus (npv) infectious dna and determinations of transfection effectiveness was studied. honeycomb moth larvae were shown to be a sensitive system for npv dna titration. deae-dextran used as a protector increased npv dna infectivity 1000-fold, ld50 in this instance being 2 x 10(8) molecules per larva. the method of npv dna infectivity determinations by the number of larvae with polyhedreae in the fatty tissue is ...19807001752
biochemical and biophysical properties of a mamestra brassicae multiple enveloped nuclear polyhedrosis virus.a multiply enveloped nuclear polyhedrosis virus from mamestra brassicae has been shown to be morphologically similar to other baculoviruses. the virus particles contain 13 polypeptides of which 4 are associated with the nucleocapsid. the polyhedron comprises of one major polypeptide of molecular weight 28,000. the dna has a mol. wt. of about 1.15 x 10(8), which is larger than that reported for other baculoviruses. the dna is a circular, supercoiled molecule of cg mol. fraction 0.448. dna fragmen ...19807002114
[virion polypeptides of the nuclear polyhedrosis virus of the silkworm].the nuclear polyhedrosis virus of the silkworm b. mori was obtained by dissolution of polyhedra at ph 10,5. it was shown that the virus particles contain a protease, which cleaves a few virion polypeptides during the dissolution of polyhedra under alkaline conditions. the proteolytic activity is inactivated by polyhedra treatment with phenylmethylsulfonylfluoride, diisopropylfluorophosphate, hgcl2 and at 80 degrees. sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed 18 virion pol ...19807011425
field-flow fractionation of alkali-liberated nuclear polyhedrosis virus from gypsy moth lymantria dispar linnaeus.rod-shaped viral particles of the gypsy moth nuclear polyhedrosis virus (npv) were obtained through alkaline dissolution of inclusion bodies isolated and purified from infected hosts. the liberated viral fraction contains a wide assortment of enveloped aggregated forms as well as enveloped monomers. this complex mixture was separated by means of sedimentation field-flow fractionation (sedimentation fff) to give a mass spectrum of the separated particles. from the elution pattern effective molecu ...19807014576
autographa californica nuclear polyhedrosis virus structural proteins compared from in vivo and in vitro sources.polyhedrin obtained from autographa californica nuclear polyhedrosis virus (acmnpv) was apparently not modified in terms of primary structure after passage through alternate host systems, both in vivo and in vitro, as investigated by two-dimensional, high-voltage electrophoresis of tryptic digests of this protein as well as amino acid analysis. n-terminal analyses were conducted using an improved technique which showed proline to be the n-terminal amino acid. in addition, passage of enveloped nu ...1979372131
characterization of the nuclear polyhedrosis virus dna of adoxophyes orana and of barathra brassicae. 1979373235
macromolecular structure of nuclear polyhedrosis virus genome.dna preparations from nuclear polyhedrosis virus (npv) of galleria mellonella l. (gml) were fractionated in high ionic strength neutral sucrose gradient. this procedure allowed a separation of supercoiled infectious dna molecules with contour length of 48--52 microns from infectious open ring dna molecule, and noninfectious linear dna molecules of the same size. in addition a heterogeneity of supercoiled dna molecules was detected. covalently closed dna molecules did not contain protein or ribon ...1979378185
[protein metabolic change in the tissues of aporia crataegi l. under the action of the nuclear polyhedrosis virus].changes in protein metabolism in the hemolymph, fat body, intestine, and gonads of aporia crataegi l. caterpilars under the influence of nuclear polyhedrosis virus were studied. electrophoresis in polyacrylamide gel was used to study acid and basic proteins of insect tissues normally and after virus infection. the virus infection was shown to result in sharp changes of the protein spectrum of various tissues and organs, particularly 3 days after infection. the main changes consisted in the loss ...1979380164
a study of two species of fish inoculated with spruce budworm nuclear polyhedrosis virus.preliminary studies conducted on rainbow trout (salmo gairdneri) and white suckers (castostomus commersoni) exposed to spruce budworm nuclear polyhedrosis virus revealed minor histopathologic changes in various organs of each fish which were interpreted as not being significant. however, an evaluation (mean values) of the total changes in groups of fish suggested that a relationship may exist in the suckers exposed to either purified polyhedra or virions. a more extensive and definitive study mu ...1979384027
interference with polyhedral inclusion body (pib) production in trichoplusia ni cells infected with a high passage strain of autographa californica nuclear polyhedrosis virus (npv). brief report.trichoplusia ni cells infected with a low passage (lp) strain of autographa californica nuclear polyhedrosis virus (npv) produce large numbers of polyhedral inclusion bodies (pibs). interference with pib production occurs when t. ni cells are first inoculated with a high passage (hp) strain and then challenged with the lp strain. pib production is reduced 100 fold to the level seen with hp virus infection only.1979389204
non-occluded particles of nuclear polyhedrosis virus infecting galleria mellonella l.: titration in vivo and in vitro. brief report.infectivity of non-occluded particles of nuclear polyhedrosis virus infecting galleria mellonella l. was determined with bioassay technique in vivo and with plaque assay method in vitro using scld 135 (quiot) established cell culture. a plaque-forming virus unit corresponds to about 50 id50 for galleria larvae injected into hemocoel for non-cloned and 0.1-1.0 id50 for a virus isolate cloned from an infective plaque.1979391180
[ultracytological studies of the morphogenesis and structure of the nuclear polyhedrosis virus in the silkworm moth, bombyx mori l].an electron-microscopic investigation was made on hypodermal tissues of silkworms spontaneously infested by nuclear polyhedral disease, due to low temperature stress. it was established that the polyhedral virus is formed in the viroplasma produced after viral cell infection, is included in the intimate and developing membrane singly or in fascicles containing several (up to 9) and is finally incapsulated in polyhedral protein. besides viral rods, various polymorphic forms were observed in the n ...1979392906
infection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus i. synthesis of intracellular proteins after virus infection.the replication of autographa californica nuclear polyhedrosis virus (acnpv) in spodoptera frugiperda cells in culture has been studied with different methods. the first virus-induced polypeptides (with molecular weights of 46k, 30k, 29k) in infected cells appeared at 3 hr postinfection. viral dna synthesis started at about 5 hr postinfection. by electron microscopy, intranuclear nucleocapsids were detected at 10 hr postinfection and at about the same time, the titer of intracellular infectious ...197916945842
infection of spodoptera frugiperda cells with autographa californica nuclear polyhedrosis virus ii. the viral dna and the kinetics of its replication.the kinetics of replication of autographa californica nuclear polyhedrosis virus (acnpv) dna in spodoptera frugiperda cells in culture were studied. viral dna replication started at about 5 hr postinfection, the rate of viral dna replication reached a maximum at about 18 hr postinfection and thereafter decreased. parental viral dna could be detected within the first hour postinfection in the total intracellular dna by the southern technique. there was no evidence for the occurrence of acnpv dna ...197916945843
characteristics of the dna from lymantria dispar nuclear polyhedrosis virus.the dna isolated from occluded lymantria dispar nuclear polyhedrosis virus consisted of double-stranded linear, open circular, and covalently closed molecules. the dna had a guanosine plus cytosine content (g + c) of 59-62% as estimated from both thermal denaturation and cscl density gradient centrifugation. the l. dispar host dna had a g + c content of 38-39%. comparisons of molecular weight estimations for viral dna by sedimentation on neutral sucrose gradients, electron microscopy, and reasso ...197918627902
physical map of the dna genome of autographa californica nuclear polyhedrosis virus.a physical map of the 88 x 10(6) dalton, circular dna genome of autographa californica nuclear polyhedrosis virus was constructed. the complete order of bamhi and xmai restriction enzyme sites was determined. the ecori and hindiii fragments were partially ordered, and their general locations, relative to the bamhi and xmai maps, were determined. alterations in the restriction endonuclease fragment patterns of natural genotypic variants of a. californica nuclear polyhedrosis virus, including tric ...197916789175
restriction maps of five autographa californica mnpv variants, trichoplusia ni mnpv, and galleria mellonella mnpv dnas with endonucleases smai, kpni, bamhi, saci, xhoi, and ecori.the restriction sites of autographa californica nuclear polyhedrosis virus (acmnpv) e2 dna were mapped for the endonucleases smai, kpni, bamhi, saci, xhoi, and ecori. the restriction maps of four other acmnpv variants, trichoplusia ni (tnmnpv), and galleria mellonella (gmmnpv) genomes were determined and compared to the endonuclease cleavage maps of acmnpv e2 dna. the viral structural polypeptides of acmnpv variants s3, e2, s1, m3, and r9 were the same when analyzed by polyacrylamide gel electro ...197916789179
genetic analysis of a baculovirus, autographa californica nuclear polyhedrosis virus i. isolation of temperature-sensitive mutants and assortment into complementation groups.temperature-sensitive (ts) mutants were isolated from the baculovirus autographa californica (alfalfa looper) mnpv, grown in spodoptera frugiperda (fall armyworm) cells in the presence of n-methyl-n'-nitro-n-nitrosoguanidine. of 567 plaque isolates screened, 27 were temperature sensitive (ts), representing a mutation frequency of 4.8%. ten ts mutants were studied in detail: six failed to yield nonoccluded virus at 33 degrees c (nov mutants), whereas the other four produced nonoccluded virus but ...197916789180
isolation, complementation, and initial characterization of temperature-sensitive mutants of the baculovirus autographa californica nuclear polyhedrosis virus.sixteen temperature-sensitive mutants of autographa californica nuclear polyhedrosis virus were isolated. several interesting phenotypes were observed. a large proportion of the mutants were unable to form polyhedral occlusion bodies (polyhedra) at the nonpermissive temperature (32.5 degrees c). at 32.5 degrees c, one mutant formed plaques in which the cells lacked polyhedra. another mutant type was defective in the production of progeny extracellular nonoccluded virus and produced a "plaque" co ...197916789181
comparison of solid-phase radioimmunoassays for baculoviruses.the sensitivity and cross-reaction of four solid-phase radioimmunoassays (ria) for trichoplusia ni nuclear polyhedrosis virus containing singly enveloped virions were investigated. the detection limits of each assay were as follows: indirect ria, 5 ng of dissolved polyhedron antigen; direct ria, 50 ng; indirect sandwich ria, 200 ng; and direct sandwich ria, 300 ng. the indirect and indirect sandwich rias showed considerable cross-reaction with other baculovirus antigens, but the direct and direc ...197816345305
plaque assay of nuclear polyhedrosis viruses in cell culture.the nuclear polyhedrosis virus of autographa californica has been titrated in spodoptera frugiperda cells by the plaque method, using a solid overlay which does not require either the use of modified culture medium or expensive purified agarose or the addition of culture medium as a liquid layer above the solid agarose. this assay is more sensitive than that using a viscous methyl cellulose overlay but less sensitive than the end-point dilution technique. neither trichoplusia ni nor bombyx mori ...197816345306
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