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

作　　者：洪帮兴[1] 江丽芳[1] 胡玉山[1] 方丹云[1] 郭辉玉[1] 

机构地区：[1]中山大学中山医学院微生物学教研室

出　　处：《中华微生物学和免疫学杂志》2004年第3期241-244,共4页Chinese Journal of Microbiology and Immunology

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)

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　 2Objective 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：23S RRNA 基因序列分析 细菌感染 鉴别诊断 生物信息学 基因芯片 

分 类 号：R346[医药卫生—基础医学]