华西口腔医学杂志

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变形链球菌临床株质子移位膜ATP酶γ亚基基因uncG遗传多态性及mRNA表达水平的研究

杨德琴1, 刘天佳2, 亓庆国3, 庄姮2, 李颂4   

  1. 1.口腔生物医学工程教育部重点实验室, 四川大学; 2.四川大学华西口腔医院牙体牙髓科, 四川成都610041; 3.山东大学口腔医院牙体牙髓科, 山东济南250012; 4.安徽医科大学口腔医院牙体牙髓科, 安徽合肥230000
  • 收稿日期:2007-02-25 修回日期:2007-02-25 出版日期:2007-02-20 发布日期:2007-02-20
  • 通讯作者: 刘天佳，Tel：028- 85501439
  • 作者简介:杨德琴（1972-），女，广西人，副教授，博士，现在遵义医学院附属口腔医院口腔内科工作
  • 基金资助:

    国家自然科学基金资助项目（30171013）；贵州省自然科学基金资助项目[黔科合计（2004）3055]；贵州省优秀科技人才省长专项基金资助项目[（2005）244]

Genetic Diver sity and mRNA Expr ession of F- ATPase Subunit uncG Gene of Streptococcus mutans Clinical Isolates

YANG De - qin1, LIU Tian- jia2, QI Qing- guo3, ZHUANG Heng2, LI Song4   

  1. 1. Key. Laboratory of Oral Biomedical Engineering of Ministry of Education, Sichuan University, Chengdu 610041, China; 2. Dept. of Operative Dentistry and Endodontics, West China College of Stomatology, Sichuan University, Chengdu 610041, China; 3. Dept. of Operative Dentistry and Endodontics, College of Stomatology, Shandong University, Jinan 250012, China; 4. Dept. of Operative Dentistry and Endodontics, College of Stomatology, Anhui University of Medical Science, Hefei 230000, China
  • Received:2007-02-25 Revised:2007-02-25 Online:2007-02-20 Published:2007-02-20
  • Contact: LIU Tian- jia，Tel：028- 85501439

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摘要：

目的研究变形链球菌临床分离株质子移位膜ATP酶（F- ATPase）的γ亚基基因uncG的遗传多态性和mRNA表达水平。方法选取在不同龋敏感人群中分离的18株高耐酸和20株低耐酸变形链球菌为实验株，扩增uncG基因，行限制性内切酶长度多态性（RFLP）分析和测序比较，应用RT- PCR法和凝胶成像系统定量软件对20株不同基因型和不同耐酸力变形链球菌uncG基因的mRNA表达水平进行评价和比较。结果ALuⅠ- RFLP和BsrⅠ-RFLP产生A、B、C、D 4种基因型，但4种基因型在不同耐酸力菌株的分布差异没有统计学意义（P>0.05）。不同基因型及不同耐酸力菌株uncG的mRNA表达水平不同，但其差异也没有统计学意义（P>0.05）。结论变形链球菌F- ATPase γ亚基基因uncG的RFLP基因型和mRNA表达水平具有明显遗传异质性，但对菌株耐酸力没有明显影响。

关键词: 变形链球菌, 质子移位膜ATP酶, 耐酸性, 基因多态性

Abstract:

Objective To study the genetic diversity and the gene expression of membrane- bound proton- translocating ATPase（F- ATPase） subunit gene uncG derived from Streptococcus mutans（S.mutans） clinical isolates. Methods 38 S.mutans strains derived from caries- active and caries- free individuals including 18 strains displaying high acid tolerance and 20 strains displaying low acid tolerance. Gene uncG was amplified with specific primers from S.mutans genomic DNA, then the PCR product was analyzed by RFLP and sequenced. The relative expression quantity of uncG gene against the housekeeping gene recA was determined by using RT- PCR method. A gel documentation system and QUANTITY ONES software were used to analyze the data results. Results It was testified that four genotypes A, B, C and D of PCR- RFLP were revealed when respectively digested with AluⅠ and BsrⅠ , but the distributions of the four genotype strains showed no difference（P >0.05）. The differences of uncG gene transcript quantities derived from different genotype or different aciduranc strains had no significance（P>0.05）. Conclusion This study indicated that uncG gene of F- ATPase obviously displayed genetic diversity and existed polymorphism at mRNA expression level, while the AluⅠ- RFLP genotypes and the expression levels would not be responsive to different acid tolerance of S.mutans strains.

Key words: Streptococcus mutans, membrane- bound proton- translocating ATPase, acid tolerance, genetic diversity

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