牙龈卟啉单胞菌PG2206基因突变株的构建

Construction of a mutant in the PG2206 gene of Porphyromonas gingivalis

目的探讨牙龈卟啉单胞菌(P.gingivalis)PG2206基因缺失突变株的构建方法,为进一步研究该基因的功能提供实验基础。方法将具有四环素抗性的重组基因载体转导入P.gingivalis W83的感受态细胞中替换PG2206,在四环素抗性的培养基上筛选阳性克隆株,命名为PG2206基因突变株。结果对阳性克隆株进行验证,成功构建出P.gingivalis W83的PG2206基因缺失的突变株。结论采用同源重组的方法可成功构建PG2206基因突变株。

Objective To determine the function of PG2206 gene from Porphyromonas gingivalis （P.gingivaIis） W83 strain, a mutant in the PG2206 gene was constructed by homologous recombination. Methods The recombinant gene with tetracycline resistance was transduced into competent cells of P.gingivalis W83. After electroporated and selected on the tetracycline brain heart infusions plates, the single colony was collected and designated as PG2206 gene-deficient mutant. Results The positive clones were verified by PCR, which indicates that the PG2206 gene was knocked out successfully. Conclusion A PG2206 gene-defective mutant was created successfully.