变形链球菌F-ATP酶基因重组质粒的构建和初步分析

Construction and initial study of recombinant plasmid containing S.mutans F-ATPase gene

目的:构建用于转化变形链球菌,含有F-ATP酶基因的重组质粒。方法:采用PCR方法,以变形链球菌基因组DNA为模板,扩增F-ATP酶β亚基5′末端序列,将克隆片段与载体pVA891酶切后连接,形成重组质粒,并对变形链球菌的转化作了初步分析。结果:构建的重组质粒经PCR鉴定、酶切鉴定和DNA序列测定,显示插入的目的片段序列正确。结论:变形链球菌目的片段与穿梭载体重组后能有效克隆,为通过同源重组特异突变变形链球菌染色体基因奠定基础。

AIM：To construct a homologous recombinant plasmid which was expected to be transformed of S. rnutans. METHODS：A region at the 5′terminus of the S. rnutans F -ATPase β subunit gene was amplified by PCR. The PCR product was inserted into vector pVA891 ,yielding recombinant plasmid. The plasmid was attempted to transform S. rnutans. RESULTS：The DNA sequence of the recombinant plasmid was identified correct in whole by PCR, restriction endonuclease and DNA sequence techniques. CONCLUSION：The recombinant plasmid of S. mutans DNA was cloned effectively. It may assist in construction of homologues recombinant mutant.