鸭Ⅰ型肝炎病毒VP1基因片段的克隆与表达载体的构建

Abstract

Abstract:

To study the expression of the duck hepatitis virus 1(DHV1) VP1 gene in E.coli ,one outer primer pair and one inter primer pair containing EcoR Ⅰ and Sal Ⅰ according to complete genome of DHV1 were designed. PCR products were re cycled by outer and inter primer pair .The PCR products and pET-32a vector were digested by EcoR Ⅰ and Sal Ⅰ to construct the recombinant plasmids.Then the recombinant plasmids were transfected into E.coli BL21 and Rosseta. After digestion, PCR and sequencing the target gene were insert expression vector correctly. With different concentrations of IPTG induced bacteria express VP1 gene, the bacterias were collected and examined by SDS-PAGE.The results showed that the VP1 gene fragments hard to express in E.coli Rosseta and BL21.

Key words: duck hepatitis virus R85952 strain; VP1 gene; cloning; expression

ZHAO Rui-hong;ZHANG Dan-jun;PAN Xiao-cheng;CHENG Bao-yan;HU Xiao-miao. Cloning of VP1 Gene Fragments of Duck Hepatitis Virus TypeⅠand Construction of its Expression Plasmid[J]. , 2010, 37(7): 67-69.

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Cloning of VP1 Gene Fragments of Duck Hepatitis Virus TypeⅠand Construction of its Expression Plasmid

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