中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (7): 1231-1234.doi: 10.3969/j.issn.1673-8225.2010.07.021

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

变形链球菌thrS基因生物信息学分析及同源重组质粒的构建

刘筱娣1，段  劲2，郭丽宏1   

  1. 1 北京大学口腔医学院•口腔医院，北京市 100081；2 湖州师范学院，浙江省湖州市  313000
  • 出版日期:2010-02-12 发布日期:2010-02-12
  • 通讯作者: 郭丽宏，博士，副教授，北京大学口腔医学院，北京市 100081 guo_lihong2008@yahoo.com.cn
  • 作者简介:刘筱娣★，女，1981年生，北京市人，汉族，2006年北京大学医学部口腔医学专业毕业，硕士，助理研究员，主要从事变形链球菌功能基因方面的研究。 kqlxd@163.com
  • 基金资助:

    北京市自然科学基金项目(7042035)。

Bioinformatic analysis of streptococcus mutans thrS gene and construction of homologous recombinant plasmids

Liu Xiao-di1, Duan Jin2, Guo Li-hong1   

  1. 1 Peking University School & Hospital of Stomatology, Beijing  100081, China; 2 Huzhou Teachers College, Huzhou  313000,  Zhejiang Province, China
  • Online:2010-02-12 Published:2010-02-12
  • Contact: Guo Li-hong, Doctor, Associate professor, Peking University School & Hospital of Stomatology, Beijing 100081, China guo_lihong2008@yahoo.com.cn
  • About author:Liu Xiao-di★, Master, Research assistant, Peking University School & Hospital of Stomatology, Beijing 100081, China kqlxd@163.com
  • Supported by:

    the Natural Science Foundation of Beijing, No. 7042035*

PDF

509

被引次数

3

摘要：

背景：变形链球菌是公认的主要致龋菌，细菌的黏附、生物膜形成、产酸、耐酸等各个表型都与细菌的致龋性相关。有研究结果提示，苏氨酰-tRNA合成酶基因(threonyl-tRNA synthetase, thrS)可能与变形链球菌的黏附、产酸、耐酸等致龋性表型相关。
目的：对苏氨酰-tRNA合成酶基因进行保守性分析，构建用于变形链球菌苏氨酰-tRNA合成酶基因敲除的重组质粒。
方法：首先通过生物信息学结合Southern Blot对苏氨酰-tRNA合成酶基因的保守性进行分析，再将变形链球菌苏氨酰-tRNA合成酶基因上下游序列分别克隆至自杀质粒pFW5上的多克隆位点，构建用于基因敲除的重组质粒。观察苏氨酰-tRNA合成酶基因的保守性及重组质粒的构建结果。
结果与结论：苏氨酰-tRNA合成酶基因在实验中所列6种变形链球菌中保守存在；经过聚合酶链反应和酶切分析，重组质粒所插入片断无误。表明重组质粒pFW5-thrS构建成功。重组质粒pFW5-thrS的构建可用于今后变形链球菌苏氨酰-tRNA合成酶基因功能的研究。

关键词: 变形链球菌, thrS, 重组质粒, 生物信息学, 基因

Abstract:

BACKGROUND: Streptococcus mutans (S. mutans) is generally considered to be the principal aetiological agent for dental caries. thrS gene may relate to the virulence of S. mutans involved in the adherence, acidogenicity and acidodurance.
OBJECTIVE: To investigate the conservation status of the thrS gene of S. mutans and to construct the homologous recombinant plasmid.
METHODS: Southern Blot was used to analyze the distribution of thrS gene in S. mutans. The upstream and downstream sequences of thrS gene were cloned respectively into multiple cloning sites of suicide plasmid pFW5 to construct the recombinant plasmid.
RESULTS AND CONCLUSION: The thrS gene was conserved in 6 strains of S. mutans in this test. By PCR analysis and enzyme digesting, it was confirmed that S. mutans thrS gene homologous recombinant plasmid was successfully constructed, which can be used in future research of construction of thrS -negative mutans of S. mutans strain UA159

中图分类号: