传染性喉气管炎病毒王岗株gB基因gC基因和gD基因的真核表达

Eukaryotic expression of gB, gC and gD genes of ILTV WG strain

将传染性喉气管炎病毒(ILTV)王岗(WG)株的gB、gC、gD基因分别克隆到真核表达载体pCAGGS的EcoRⅠ位点,经过酶切、测序分析,筛选鉴定出含有gB、gC、gD基因的重组质粒,分别命名为pCAGGgB、pCAGGgC、pCAGGgD。用质粒纯化试剂盒对重组质粒进行了纯化,将纯化后的质粒转染293T细胞,用间接免疫荧光技术检测了目的蛋白的表达情况。检测结果表明,目的蛋白得到了真实的表达。

The genes gB, gC and gD amplified from infectious laryngotracheitis virus (ILTV) Chinese Wanggang (WG) strain were cloned into the EcoRⅠ site of the eukaryotic expression vector pCAGGS, respectively. The positive recombinant plasmids were screened by transforming into E.coli DH5α cells, which were named pCAGG-gB, pCAGG-gC and pCAGG-gD respectively after identification by restriction endonuclease analysis and sequencing analysis. Then the recombinant plasmids were amplified and purified and transfected into 293T cells in vitro. The indirect immunofluorescence analysis was used to detect the expression of target proteins. The result showed that the target proteins had been successfully expressed.