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Partial Nucleotide Sequence And Genomic Organization Of In Korean Isolate Of Helicoverpa Armigera Singlenucleocapsid Nucleopolyhedrosis Virus

Posted on:2004-01-15Degree:DoctorType:Dissertation
Country:ChinaCandidate:G R ZhaoFull Text:PDF
GTID:1103360125955707Subject:Microbiology
Abstract/Summary:PDF Full Text Request
Helicovera armigera nucleopolyhedrosis(HaNPV) is a member of the family Baculovirus, is designated as single (S) or multiple (M) depending on the potential number of nucleocapsid packaged in a virion.H. armigera single nucleocapsid nucleopolyhedrosis(HaSNPV) were isolated in differente countries, they are G4, HaS-BW, HaS-ZN, SHANGHAI isolate and HaS-SA.Due to its high virulence, HaSNPV has been adopted for mass production as a viral pesticide and has been widely used to control the insect pests in China and in other countries.Biochemical, serological and molecular studies revealed that the various SNPVs have a high degree of similarity. But Helicovera SNPVs containg different genotypes share, show differences in virulence and rstiction enzyme profiles.HaSNPV-KO was studied on the shape and structure, structural polypeptide, restriction pattern.Diameter of polyhedron is 0.7-2.0μm, size of virion is 326nm.69nm. Assayed by SDS-PAGE, polyhedron of HaSNPV-KO contained a single polypeptide whose molecular is 28.7kDa, genome is 130.18kb. Genome digested by BamHI, EcoRI, HindIII and PstI, map attained is very similar to the other isolates reported.Insight into the molecular characteristics that specify the biological properties of HaSNPV is important not only to understand the basis of host range and virulence, but also for the successful and bio-safe genetic modification of this virus. Despite the extensive usage of HaSNPV in pest control over many decades, the molecular knowledge of the HaSNPV genome is limited to viral DNA restrictionenzymes profile analysis.The nucleotide sequence of G4 isolate(Gene Bank 271059), HzSNPV(Gene Bank 334030) and SHANGHAI isolate(Gene Bank 303045) have been determined.The genome of G4 was assembled into a conyiguous sequence of 131403bp.The size are very similar to the HzSNPV(130869bp) and SHANGHAI isolate(130760bp).In order to investigate the genomic organization of the HaSNPV-KO, determind BamHI-I fragment, BamHI-H fragment and BamHI-G fragment, compared with the other isolates.The BamHI-I fragment contained p26 gene, pl0 gene and 3'termini of p74 gene; the BamHI-H fragment contained gp37 gene,hr2 homologous region and 5'termini of bro-b gene; the 3'termini of bro-b gene, he65 gene,hr3 homopogous region, iap2 gene and 3'termini of HaORF63 gene were localized BamHI-G fragment of genome.Nucleotide sequence aligment indicated that these gene has an extremely high degree of indentity with that of other isolate.hr2 homologous region have 1150bp,which is equal with the G4 isolate. And hr3 homologous region have 75 9bp,which is equal with the G4 isolate.The secondary protein structure prediction and hydrophobic amino acid of the p26 gene, pl0 gene, gp37 gene, bro-b gene, he65 gene and iap2 gene were performed with computer programs.
Keywords/Search Tags:HaSNPV-KO, structural polypeptide, restriction map, nucleotide sequence, genomic organization
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