Ⅰ型鸭肝炎病毒VP3基因的原核表达及其抗原性

›› 2011, Vol. 23 ›› Issue (1): 66-69.

Ⅰ型鸭肝炎病毒VP3基因的原核表达及其抗原性

刘燕,庞安娜,韦强,鲍国连*,季权安,肖琛闻

浙江省农业科学院畜牧兽医研究所, 浙江杭州 310021

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-01-25 发布日期:2011-01-25

Prokaryotic expression and antigenicity analysis of VP3 gene of Duck hepatitis virus Ⅰ

LIU Yan;PANG An\|na;WEI Qiang;BAO Guo\|lian*;JI Quan\|an;XIAO Chen\|wen

Institute of Animal Husbandry and Veterinary, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)

Received:1900-01-01 Revised:1900-01-01 Online:2011-01-25 Published:2011-01-25

摘要/Abstract

摘要： 参考GenBank 中的Ⅰ型鸭肝炎病毒(DHV Ⅰ)全基因序列设计引物,采用RT-PCR方法扩增了DHV Ⅰ VP3 基因,将VP3 基因与pET-28a(+)表达载体连接后, 转化宿主BL21(DE3), 用01 mmol·L-1的IPTG诱导表达, 收集菌液进行SDS-PAGE电泳, Western-blotting分析蛋白免疫原性。电泳结果表明, VP3在大肠杆菌中表达量较高, 表达产物的分子量约为27 kD。免疫印迹试验表明 Ⅰ型鸭肝炎病毒VP3蛋白在大肠杆菌中表达产物具有免疫原性。

关键词: Ⅰ型鸭肝炎病毒, VP3基因, 克隆表达

Abstract: In this study, a pair of special primers,according to the complete genome of Duck hepatitis virus Ⅰ(DHV-Ⅰ) , was designed to amplify VP3 gene of DHV-Ⅰ. VP3 was subcloned into pET-28a(+), then the recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and induced by 0.1 mmol·L-1 IPTG. The bacteria containing pET-28a(+)VP3 were collected and examined by SDS-PAGE and Western-blotting. The results showed that VP3 protein was well expressed in E. coli. The molecular weight of the recombinant protein was 27 kD. The protein could be recognized by the specific antibody against DHV-Ⅰ.

Key words: Duck hepatitis virusⅠ, VP3 gene, cloning and expression

刘燕;庞安娜;韦强;鲍国连*;季权安;肖琛闻. Ⅰ型鸭肝炎病毒VP3基因的原核表达及其抗原性[J]. , 2011, 23(1): 66-69.

LIU Yan;PANG An\|na;WEI Qiang;BAO Guo\|lian*;JI Quan\|an;XIAO Chen\|wen. Prokaryotic expression and antigenicity analysis of VP3 gene of Duck hepatitis virus Ⅰ[J]. , 2011, 23(1): 66-69.

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