上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (5): 684-.doi: 10.3969/j.issn.1674-8115.2011.05.034

• 技术与方法 • 上一篇    下一篇

不同16S rDNA通用引物对DGGE进行牙周微生物群落分析的影响

周彦玢, 束 蓉, 刘大力   

  1. 上海交通大学 医学院附属第九人民医院牙周病科 |上海市口腔医学重点实验室, 上海 200011
  • 出版日期:2011-05-28 发布日期:2011-05-27
  • 通讯作者: 束 蓉, 电子信箱: shurong123@hotmail.com。
  • 作者简介:周彦玢(1986—), 女, 博士生；电子信箱: kathleen12@163.com。
  • 基金资助:

    “十一五”国家科技支撑计划（2007BAI18B02）；上海市重点学科建设项目（S30206）；上海市教委科研创新项目（09YZ78）

Effect of different 16S rDNA universal primers on DGGE patterns of periodontal microbial community

ZHOU Yan-bin, SHU Rong, LIU Da-li   

  1. Department of Periodontology, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Online:2011-05-28 Published:2011-05-27
  • Supported by:

    National Science and Technology Foundation of the Eleventh Tenth-Five Year Plan, 2007BAI18B02；Shanghai Key Discipline Construction Program, S30206；Shanghai Education Committee Foundation, 09YZ78

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摘要：

目的 初步评价不同的16S rDNA片段对变性梯度凝胶电泳（DGGE）进行牙周微生物群落分析结果的影响。方法 选取3种牙周主要可疑致病菌的标准菌株，通过构建质粒DNA分别获得16S rDNA V3区、V3~V5区和V6~V8区片段，测序鉴定并进行BLAST验证；将测序正确的质粒DNA经DGGE分离和硝酸银染色后获得条带图谱。结果 由3对通用引物扩增得到的DGGE图谱并不相同。V3区引物获得的并非单一条带，且不同菌株间没有识别度；V3~V5区和V6~V8区的图谱显示：3种菌株的DNA片段均得到了单一条带，且区分性较好。结论 16S rDNA V3~V5区和V6~V8区的两对引物可用于龈下菌群多样性的研究。

关键词: 龈下菌群, 16s rDNA, 变性梯度凝胶电泳, 可疑致病菌

Abstract:

Objective To preliminarily assess the effect of different 16S rDNA fragments on denaturing gradient gel electrophoresis (DGGE) patterns of periodontal microbial community. Methods Three typical periodontal bacterial strains were selected, plasmid DNA was constructed to get 16S rDNA V3, V3-V5 and V6-V8 region fragments, and sequencing analysis and BLAST identification were performed. The bands of strains were obtained after DGGE isolation and silver nitrate staining of sequenced plasmid DNA. Results DGGE profiles by three universal primers were different. The bands of 16S rDNA V3 region fragments of typical strains were not single, and bands of different strains could not differ from each other. The single band of 16S rDNA V3-V5 and V6-V8 region fragments of each typical strain was found. Conclusion 16S rDNA V3-V5 and V6-V8 region fragments are suitable for periodontal bacteria structure analysis.

Key words: subgingival bacteria community, 16s rDNA, denaturing gradient gel electrophoresis, putative pathogen