PCR-SSCP技术在嗜盐放线菌链单孢菌属快速筛选中的应用

Rapid Selection of Halophilic Streptomonospora Strains by PCR-SSCP

摘要:

为提高嗜盐放线菌的研究效率, 快速、准确的从大量分离菌株中去除重复菌株、筛选出目的菌株, 在特异性引物快速定属的基础上, 以嗜盐放线菌链单孢菌属的34株菌株为研究对象, 采用与PCR相结合的单链构象多态性分析(PCR-SSCP), 扩增出16S rRNA基因中的两个高变区, 根据结果对34株菌株进行聚类分析, 并将其16S rRNA基因片段测序予以验证。结果表明, 聚类后34株菌株可大致分为3类, 且与16S rRNA基因片段分析结果一致。从而可快速去除重复菌株并反映出菌株间的系统进化关系。同时实验数据可构建成库, 使后续分离菌株的筛选工作只需比对数据即可完成, 利于提高工作效率, 降低实验成本。

Abstract:

To improve the efficiency of halophilic actinobacteria screening and carry out the rapid selection of targeted strains, we tested 34 strains of Streptomonospora by PCR-single strand conformation polymorphism analysis (PCR-SSCP) based on genus-specific primers for the PCR identification. This approach employs PCR with two pairs of primers located in the 16S rRNA sequence flanking two variable region, then build clustering tree according as SSCP data. Synchronously, we sequenced all the 16S rRNA partial sequences for these strains to verify them. The results showed that the PCR-SSCP analysis was an efficient, easy-to-handle and economic method for rapid selection of halophilic actinobacteria resources.