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Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome
Wang, HZ; Deng, F; Pijlman, GP; Chen, XW; Sun, XL; Vlak, JM; Hu, ZH; Hu, ZH, Chinese Acad Sci, Wuhan Inst Virol, Joint Lab Invertebrate Virol, Wuhan 430071, Peoples R China
2003
Source PublicationVIRUS RESEARCH
Volume97Issue:2Pages:57-63
AbstractPurification of genotypes from baculovirus isolates provides understanding of the diversity of baculoviruses and may lead to the development of better pesticides. Here, we report the cloning of different genotypes from an isolate of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV) by using a bacterial artificial chromosome (BAC). A transfer vector (pHZB10) was constructed which contained an Escherichia coli mini-F replicon cassette within the upstream and downstream arms of HaSNPV polyhedrin gene. Hz2e5 cells were co-transfected with wild-type HaSNPV DNA and pHZB10 to generate recombinant viruses by homologous recombination. The DNA of budded viruses (BVs) was used to transform E. coli. One of the bacmid colonies, HaBacHZ8, has restriction enzyme digestion profiles similar to an in vivo cloned strain HaSNPV-G4, the genome of which has been completely sequenced. For testing the oral infectivity, the polyhedrin gene of HaSNPV was reintroduced into HaBacHZ8 to generate the recombinant bacmid HaBacDF6. The results of one-step growth curves, electron microscopic examination, protein expression analysis and bioassays indicated that HaBacDF6 replicated as well as HaSNPV-G4 in vitro and in vivo. The biologically functional HaSNPV bacmids obtained in this research will facilitate future studies on the function genomics and genetic modification of HaSNPV. (C) 2003 Elsevier B.V. All rights reserved.
KeywordBaculoviruses Genotypes Hasnpv Bacmid Polyhedrin Gene
DepartmentChinese Acad Sci, Wuhan Inst Virol, Joint Lab Invertebrate Virol, Wuhan 430071, Peoples R China; Chinese Acad Sci, Wuhan Inst Virol, Key Lab Mol Virol, Wuhan 430071, Peoples R China; Chinese Acad Sci, Wuhan Inst Hydrobiol, Wuhan 430072, Peoples R China; Univ Wageningen & Res Ctr, Virol Lab, NL-6709 PD Wageningen, Netherlands
Subject AreaVirology
Indexed BySCI
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9598
Collection期刊论文
Corresponding AuthorHu, ZH, Chinese Acad Sci, Wuhan Inst Virol, Joint Lab Invertebrate Virol, Wuhan 430071, Peoples R China
Recommended Citation
GB/T 7714
Wang, HZ,Deng, F,Pijlman, GP,et al. Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome[J]. VIRUS RESEARCH,2003,97(2):57-63.
APA Wang, HZ.,Deng, F.,Pijlman, GP.,Chen, XW.,Sun, XL.,...&Hu, ZH, Chinese Acad Sci, Wuhan Inst Virol, Joint Lab Invertebrate Virol, Wuhan 430071, Peoples R China.(2003).Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome.VIRUS RESEARCH,97(2),57-63.
MLA Wang, HZ,et al."Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome".VIRUS RESEARCH 97.2(2003):57-63.
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