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23S rRNA基因序列分析及其在细菌鉴别诊断中的应用

Sequence analysis of 23S rRNA gene and its amplification in bacteria discriminative identification

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【作者】 洪帮兴江丽芳胡玉山方丹云郭辉玉

【Author】 HONG Bang xing, JIANG Li fang, HU Yu shan, FANG Dan yun, GUO Hui yu. Department of Microbiology, School of Medicine, Sun Yatsen University, Guangzhou 510089, China

【机构】 中山大学中山医学院微生物学教研室中山大学中山医学院微生物学教研室 510089广州现工作单位:518033深圳市福田区疾病预防控制中心510089广州

【摘要】 目的 研究细菌 2 3SrRNA基因序列的差异 ,为细菌鉴别诊断和相关研究提供科学依据。方法 设计合适的通用引物、寻找恰当的扩增条件扩增常见细菌的 2 3SrRNA基因片段。通过序列测定和运用生物信息学分析不同种属细菌 2 3SrRNA基因的保守序列、变异规律及菌株间种系进化关系。结果 扩增出常见引起食源性感染的 16属 (种 )细菌 2 3SrRNA基因片段。部分扩增产物进行了限制性酶切鉴定和序列测定。序列比较研究获得 2 1个 2 3SrRNA的通用保守序列 ,2 3SrRNA基因保守序列与变异区在种系间呈间隔分布 ,大量变异区呈“马赛克”式分布。种系间进化关系与 16SrRNA分析结果一致。结论  2 3SrRNA基因序列的分布规律为进行细菌的鉴别诊断及基因芯片的研制提供了重要的理论和实验基础

【Abstract】 Objective To study the difference of bacteria 23S rRNA gene and provide a tool for bacteria discriminative identification. Methods The 23S rRNA gene fragment of different bacteria was amplified. The amplification fragments were identified with restriction enzymes analysis and sequencing. With bio soft and the GenBank data, the 23S rRNA conservative regions, mutant regions and their relation to the genetic development of bacterial strains were studied. Results 23S rRNA gene fragment of 16 genera(species) bacteria was amplified and identified with restriction enzymes HindⅢ, KpnⅠ and SalⅠ. The 23S rRNA gene fragment of 6 species were sequenced. 21 conservative regions were obtained. Conservative and mutant regions were separately distributed and most small mutant regions distributed like morsac among conservative regions. The phylogenetic trees among different bacteria drawn from the full lenth 23S rRNA gene or partial fragment were supported by results from 16S rRNA. Conclusion The sequence distribution of 23S rRNA gene would be a solid base for the discriminative identification of bacteria strains and the construction of diagnosis gene chip.

【关键词】 23SrRNA同源性遗传学
【Key words】 S rRNAHomologyGenetics
【基金】 广东省自然科学基金资助项目 ( 5 3 2 0 14 2 0 2 0 2 85 0 111)
  • 【文献出处】 中华微生物学和免疫学杂志 ,Chinese Journal of Microbiology and Immunology , 编辑部邮箱 ,2004年03期
  • 【分类号】R346
  • 【被引频次】43
  • 【下载频次】390
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