变形链球菌表面蛋白PAc功能区基因重组质粒的亚克隆构建

Subcloning Construction of a Recombinant with Functional Regions of PAc Protein Encoding Gene of S. mutans

将变形链球菌 pac基因待表达部分克隆入哺乳动物表达质粒中构建成将用作 DNA免疫防龋研究的重组质粒。用 PCR扩增出 pac基因的 2个高度保守区域 ,扩增产物以 p GEM- T为载体进行亚克隆后 ,再克隆至高效哺乳动物表达质粒 p CI构建成重组体 ,重组质粒 p CIA- P经限制性酶切分析及 DNA测序分析证实克隆构建正确。对含 pac基因重要抗原决定簇区域的成功克隆为 DNA免疫防龋研究完成了重要准备。

The expressed DNA fragment of pac gene was cloned into a mammalian expression plasmid to construct a recombinant that could be used as anti-caries DNA vaccine in animal afterwards. Two highly-conservative regions of pac gene were amplified. The PCR product was then subcloned into pGEM-T plasmid and was finally cloned into pCI, a high-efficiency mammalian expression plasmid. The recombinant pCIA-P was verified to contain desired insert after digestion of restriction enzymes and sequencing analysis of DNA. The successful cloning of important antigenic determinant-containing regions of pac gene is a significant preparation for the research of anti-caries DNA vaccination.