PCR快速鉴定actinobacteria三种模板制备方法的比较  

Comparasion of three PCR template preparation methods for rapid identification of actinobacteria

作　　者：徐平[1] 李文均[1] 徐丽华[1] 姜成林[1] 

机构地区：[1]教育部微生物资源开放研究重点实验室云南大学省微生物研究所

出　　处：《中国抗生素杂志》2003年第7期388-390,396,共4页Chinese Journal of Antibiotics

基　　金：国家科技部基础研究重大项目 (No .2 0 0 1CCC0 0 60 0 ) ;云南省自然科学基金项目 (No .2 0 0 1C0 0 0 1Q) ;云南省教育厅基金项目 (No.0 1 1 1 1 1 34;No.0 2QJ0 77)

摘　　要：目的　本研究旨在建立准确、简便、快速的放线细菌鉴定技术 ,为普通和极端环境放线细菌资源的调查和开发利用创造条件。方法　从放线细菌固体培养基上挑取少量菌体 ,用微波炉法快速制备基因组DNA作为PCR模板 ,与液体培养法得到的菌体以超声波法或冻融法制备的模板进行了PCR扩增效果的比较研究。结果　PCR检测结果表明微波炉法制备的模板可进行有效的体外扩增 ,目的条带特异 ,而超声波法或冻融法并不对所有菌株有效 ,并有非特异扩增产物产生。结论　组合微波炉法快速制备放线细菌基因组DNA技术和 2 3SrRNA特异插入序列PCR扩增技术建立了准确、简便、快速的actinobacteria鉴别体系。Objective A rapid, specific and convenient method for distinguishing actinobacteria from bacteria was proposed. Method Genomic DNA was prepared by microwave based DNA extraction method; the ultrasonic based DNA extraction method and the freeze thaw based DNA extraction method respectively. The 23S rRNA gene fragment was amplified with these templates for identification of actinobacteria. Result The result showed that the effective and specific amplification could be done with the DNA derived from microwave based DNA extraction method. Whereas the DNA derived from the other two methods could not be validly used. Conclusion The method combined with microwave based DNA extraction method and 23S rRNA gene insert fragment PCR amplification could be used to identify actinobacteria from bacteria specifically, rapidly and conveniently.

关 键 词：ACTINOBACTERIA 微波炉法 超声波法 冻融法 基因组DNA提取 放线细菌 鉴定技术 

分 类 号：Q93－331[生物学—微生物学]