Ⅰ型鸭肝炎病毒VP1基因的克隆和表达  

Cloning and Expression of VP1 Gene of Duck Hepatitis Virus 1

作　　者：吕敏娜[1] 戚南山[1] 覃宗华 廖申权[1] 余劲术[1] 袁建丰[1] 吴彩艳[1] 孙铭飞[1] 

机构地区：[1]广东省农业科学院兽医研究所,广东广州510640 [2]广州英赛特生物技术有限公司,广东广州510663

出　　处：《动物医学进展》2012年第2期13-16,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30471300);NSFC-广东省联合基金项目(U0831004);广东省国际科技合作专项(2008A050200015);广东省自然科学基金项目(1045106001006126);广东省农业科学院院长基金项目(201014)

摘　　要：通过RT-PCR方法克隆出GD株Ⅰ型鸭肝炎病毒(DHV-1)的结构蛋白VP1基因片段,VP1基因片段和原核表达载体pMAL-C2X均以EcoRⅠ、HindⅢ双酶切后构建获得重组质粒pMAL-C2X-VP1,经限制性酶切和序列测序证明,目的基因正确插入表达载体,转入E.coli BL21(DE3),构建重组表达菌E.coliBL21/pMAL-C2X-VP1,经IPTG诱导,SDS-PAGE检测结果表明,DHV-1的结构蛋白VP1能在E.coliBL21(DE3)中表达,获得可溶性重组蛋白,表达产物的分子质量约为69ku。A pair of primers were designed and synthesized to amplify VP1 gene by RT-PCR from total RNA isolated from the duck hepatitis virus 1（Guangdong strain,GD）.The confirmed amplicon product by sequencing was digested with EcoR Ⅰand Hind Ⅲ,and ligated into pMAL-C2X vector for transformation into bacterial competent cells.Several clones were sequenced and one of the clones containing correct sequence was transformed into E.coli Rosetta（DE3） cells and induced with IPTG.SDS-PAGE analysis showed that the molecular weight of fusion protein MBP-VP1 was 69 ku.

关 键 词：Ⅰ型鸭肝炎病毒 VP1基因 重组表达 

分 类 号：S852.659.6[农业科学—基础兽医学] S858.32[农业科学—兽医学]