第一类整合酶表达载体的构建及鉴定

Construction and Indentification of the Class1 Integrons Expression Vector

摘要:

Inti1基因是编码整合酶的基因，采用PCR方法对Inti1基因片断进行扩增，扩增子采用Ned1和BamH1双酶切，并克隆到pET19b载体上。把构建好的载体转化至E.coli BL21（DE3）中，在IPTG下进行诱导表达。经SDS-PAGE电泳鉴定以及使用特异性抗His-整合酶单克隆抗体的Western-Bloting证明该重组E.coli BL21（DE3）-Inti1在IPTG诱导下可表达整合酶，此为整合酶表达调控研究奠定了基础。

Abstract:

Class1 integrase gene was amplified by PCR, then the amplicon was connected to pET19b vector after cutting by Ned1 and BamH1. The recombinant plasmids were induced to express integrase in E.coli BL21(DE3), and the integrase was identified by SDS-PAGE and Western Blot. The results showed that the integrase can be expressed in E.coli BL21(DE3).