变形链球菌耐氟菌株中耐酸相关基因gluA 突变的检测及其意义

J4 ›› 2011, Vol. 37 ›› Issue (1): 80-83.

变形链球菌耐氟菌株中耐酸相关基因gluA 突变的检测及其意义

刘莉¹, 张志民¹, 王丽颖², 郑鹏¹, 李会¹

1.吉林大学口腔医院牙体牙髓病科|吉林长春 130021；2.吉林大学白求恩医学院生物化学与分子生物学实验中心|吉林长春 130021

收稿日期:2010-11-01 出版日期:2011-01-28 发布日期:2010-01-28
通讯作者: 张志民(Tel：0431-85579300，E-mail：zhangzm1964@sina.com) E-mail:zhangzm1964@sina.com
作者简介:刘莉（1984-）|女|吉林省长春市人|在读医学硕士|主要从事牙体牙髓病基础及临床研究。
基金资助:
吉林省科技厅科研基金资助课题（200905177）

Detection of mutation of acid-resistant gene gluA in |fluoride-resistant strain of streptococcus mutans

LIU Li¹, ZHANG Zhi-Min¹, WANG Li-Ying², ZHENG Peng¹, LI Hui¹

1.Deparment of Endodontics and Operative Dentistry|School of Stomatology|Jilin University,Changchun 130021|China；2.Experiment Center of Biochemistry and Molecular Biology|Norman BethuneCollege of Medicine|Jilin University|Changchun 130021|China

Received:2010-11-01 Online:2011-01-28 Published:2010-01-28

摘要/Abstract

摘要：

目的：检测变形链球菌耐氟菌株中耐酸相关基因gluA是否发生突变，阐明变形链球菌耐氟菌株耐酸性的变化。方法：采用变形链球菌国际参考株在体外人工诱导形成变形链球菌耐氟菌株，碱裂解法提取变形链球菌耐氟菌株的基因组，根据GenBank上发表的变形链球菌不同菌株中耐酸相关基因gluA的基因序列，在保守区设计上下游引物，以变形链球菌耐氟菌株的基因组为模板进行PCR扩增，挤胶法回收PCR产物，利用pMD18-T载体进行T/A克隆，构建重组质粒，进行酶切鉴定及测序。结果：变形链球菌耐氟菌株耐酸相关基因gluA的基因序列与GenBank报告的变形链球菌耐酸相关基因gluA序列经BLASTN比对完全相同，没有碱基发生突变。结论：变形链球菌耐氟菌株中耐酸相关基因gluA未发生突变，该基因对变形链球菌耐氟菌株的耐酸性没有影响。

关键词: 变形链球菌；耐氟菌株；耐酸相关基因；gluA；重组质粒；基因突变

Abstract:

Abstract:Objective To detect the mutation of acid-resistant gene gluA in the fluoride-resistant strain of streptococcus mutans and clarify the acid-resistant change of fluoride-resistant strain. Methods Streptococcus mutans were artificially induced into fluoride-resistant strain of streptococcus mutans in vitro.The genome of fluoride- resistant strain of streptococcus mutans was obtained by alkaline method;in accordance with primer design principles,according to the published GenBank sequence of streptococcus mutans gluA, a pair of primers were designed for PCR.The PCR products were cloned into pMD18-T vector and the recombinant plasmid was constructed.The insertion fragment was identified with restrictive enzyme and sequenced. Results By BLASTN comparison,the sequence of acid-risistant gene gluA of the fluoride-resistant strain of streptococcus mutans was the same with that reported in GenBank. Conclusion The acid-resistant gene gluA of the fluoride-resistant strain of streptococcus mutans has not been any mutation which has no impact on the acid-resistant character of fluoride-resistant strain of streptococcus mutans.

Key words: streptococcus mutans；fluoride-resistant strain；acid-resistant gene gluA;recombinant plasmid;gene mutation

中图分类号:

R781.12

刘莉, 张志民, 王丽颖, 郑鹏, 李会. 变形链球菌耐氟菌株中耐酸相关基因gluA 突变的检测及其意义[J]. J4, 2011, 37(1): 80-83.

LIU Li, ZHANG Zhi-Min, WANG Li-Ying, ZHENG Peng, LI Hui. Detection of mutation of acid-resistant gene gluA in |fluoride-resistant strain of streptococcus mutans[J]. J4, 2011, 37(1): 80-83.