口腔链球菌丙酮酸氧化酶基因的克隆和序列分析

Pyruvate Oxidase Gene from Streptococcus sanguis: Molecular Cloning and Sequence Analysis of the Gene

目的 阐明血链球菌产生过氧化氢及其调节的分子机理。方法 根据已知的肺炎链球菌丙酮酸氧化酶基因 (spx B)序列设计 PCR引物 ,扩增血链球菌 ATCC10 5 5 7的丙酮酸氧化酶基因 ,以 p U C18及 M13mp18、M13m p19为载体进行克隆和亚克隆 ,并进行序列分析。结果 成功地从血链球菌 ATCC10 5 5 7扩增出丙酮酸氧化酶基因 ,获得该基因的全部序列 (1788bp) ,具有完整的开放读框 ,能编码 5 91个氨基酸的多肽。结论 血链球菌丙酮酸氧化酶基因的克隆和序列分析 。

Objective To clone and sequence the gene of pyruvate oxidase (Sopox) from Streptococcus sanguis. Methods The PCR primers for Sopox gene were designed and synthesized according to the sequence of pyruvate oxidase (spxB) gene of S.pneumonia. The amplified PCR product was cloned into pUC18 and then subcloned into M13mp18 and M13mp19. The DNA sequence of the gene was analyzed. Results Sopox gene was successfully amplified from S.sanguis ATCC10557. The nucleotide sequence of the whole gene was revealed to be 1788 base pairs with one open reading frame coding pyruvate oxidase with 591 amino acid residuals. Conclusion The clone and DNA sequence of Sopox gene were obtained which could serve as a foundation on which to elucidate the molecular mechanisms of hydrogen peroxide production and its regulation by oral streptococci.