卡介苗ERP基因置换型打靶载体的构建

Construction of the gene targeting vector of ERP gene in BCG

目的：构建ERP基因打靶载体。方法：卡介苗菌体外培养,扩增ERP基因两侧序列,连接载体与目的片段,筛选阳性克隆并鉴定。结果：PCR扩增插入片段大小与预期相符,鉴定证实PCR产物及插入片段为所需目的基因片段。结论：成功构建了用于卡介苗菌基因打靶的置换型载体,为今后构建ERP基因敲除株的卡介苗突变株的研究奠定基础。

Objective： In order to knockout erp gene in BCG,the sideward gene of erp was amplified and gene targeting vector was constructed. Methods：Culturing the BCG in vitro,extracting the genome DNA,amplifying the targeted gene with PCR,constructing the gene targeting vector and idcntitying it. Results： The fragment was amplified successfully. The replacement vector with deleted erp gene was constructed. Conclusion： Constructing successfully the replacement vector which is used to the gene knockout in BCG.