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	Efficient method to generate homologous recombinant baculovirus genomes in e-coli
	Hou, S; Chen, X; Wang, H; Tao, M; Hu, Z
刊名	Biotechniques
	2002-04-01
卷号	32期号:4页码:783-+
ISSN号	0736-6205
通讯作者	Hu, z()
英文摘要	Here we describe a convenient method to generate homologous recombinant baculoviral genomes in e. coli. the recombination takes place with the aid of recombination enzymes provided by the phage i red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. proof of concept was provided when the cathepsin gene (v-cath) of the helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (hasnpv) was replaced by the chloramphenicol resistance gene (cm-r). first, cm-r was inserted between the flanking sequences of the hasnpv v-cath. each of the flanking regions was about 1 kb. the fragment was linearized and electroporated into bacteria containing both the hasnpv bacmid and the lambda red system. recombinant bacmids resistant to chloramphenicol were selected. in comparison to the standard co-transfection/plaque assays, this method significantly reduces the time required to construct baculovirus knock-out mutants. it may also be useful in the manipulation of other large viral genomes.
WOS关键词	ESCHERICHIA-COLI ; BACTERIOPHAGE-LAMBDA ; GENES ; MUTAGENESIS ; FRAGMENTS ; INSERTION ; DELETION ; SYSTEM
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemical Research Methods ; Biochemistry & Molecular Biology
语种	英语
出版者	EATON PUBLISHING CO
WOS记录号	WOS:000174881200011
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2376106
专题	武汉病毒研究所
通讯作者	Hu, Z
作者单位	1.Chinese Acad Sci, Wuhan Inst Virol, Joint Lab Invertebrate Virol, Wuhan 430071, Hubei, Peoples R China 2.Huazhong Agr Univ, Wuhan, Hubei, Peoples R China
推荐引用方式 GB/T 7714	Hou, S,Chen, X,Wang, H,et al. Efficient method to generate homologous recombinant baculovirus genomes in e-coli[J]. Biotechniques,2002,32(4):783-+.
APA	Hou, S,Chen, X,Wang, H,Tao, M,&Hu, Z.(2002).Efficient method to generate homologous recombinant baculovirus genomes in e-coli.Biotechniques,32(4),783-+.
MLA	Hou, S,et al."Efficient method to generate homologous recombinant baculovirus genomes in e-coli".Biotechniques 32.4(2002):783-+.

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