GTF-PAc融合防龋DNA疫苗的研究(Ⅰ)携带GLU基因片段的真核表达质粒pGLU的构建

Study of GTF-PAc fusion anticaries DNA vaccine (Ⅰ):construction of eukaryotic expression plasmid pGLU carrying GLU gene fragment.

目的 :将变形链球菌葡糖基转移酶GTF I的编码葡聚糖结合区 (GLU)的序列克隆到真核载体 pCI中 ,为构建GTF PAc融合防龋DNA疫苗做准备。方法 :用PCR的方法扩增含gtfB基因的重组质粒 pYNB13的GLU区序列 ;将载体 pCI和GLU扩增片段分别酶切 ,体外连接 ,构建成真核表达质粒pGLU ,酶切电泳鉴定。 结果 :PCR获得了glu目的片段 ,质粒 pGLU含有 glu目的片段。 结论 :构建出了含有gtfB基因重要抗原决定簇区的真核表达质粒

Objective:To clone the glu fragment of Streptococcus mutans glucosyltransferase GTF I into eukaryotic expression plasmid pCI and prepare for constructing fusion GTF PAc anticaries DNA vaccine.Methods:The glu fragment was amplified from pYNB13 plasmid carrying gtfB gene;pCI vector and glu fragment was digested by restriction enzymes and connected together to get the recombinant eukaryotic expression plasmid pGLU.Results:pGLU plasmid exactly carried the glu fragment.Conclussion:The eukaryotic expression plamid pGLU including the important antigen determinant of gtfB gene had been constructed successfully. [