反义乙型肝炎病毒S基因T载体克隆的构建

Construction and identification of antisense HBV S gene cloning by T vector

目的:构建含部分乙型肝炎病毒(HBV)S基因反义序列的基因克隆。方法:根据GenBank中HBVS基因序列和计算机软件mCLONE分析的结果,设计S基因2侧引物,直接扩增出适合长度的反义HBVS基因,与PMD18T载体连接后转化DH5α,筛选阳性克隆,抽提重组质粒并进行PCR及酶切鉴定,再行序列分析。结果:获得226bp含限制性内切酶位点的阳性产物,T载体克隆经PCR及酶切鉴定和序列分析后证实,克隆片段的反向序列与GenBank中该基因的序列同源性为97%。结论:成功构建了含反义HBVS基因部分序列的T载体克隆。

Aim:To construct the PMD18T vector containing part of the antisense HBV S gene.Methods:Sense and antisense primers of HBV S gene were designed according to the sequence of the HBV S gene from GenBank and the analytical results of the mCLONE software. Using the primers,the proper length antisense HBV S gene was obtained by PCR. The amplified DNA fragments were ligated into PMD18T vector,and then transformed into DH5α.The positive clones were screened out and the recombinant plasmids were isolated from such c...