重组乳杆菌质粒拷贝数测定方法的建立  

Establishment of the detection method of the recombinant lactic acid bacteria expression vector plasmid copy number

作　　者：王宇鹏[1] 齐浩[1] 林红丽[1] 王嵩[1] 侯喜林[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国兽医杂志》2015年第9期40-42,共3页Chinese Journal of Veterinary Medicine

摘　　要：为建立一种快速、准确、特异的重组乳杆菌表达质粒拷贝数的定量检测方法,根据SYBR Green I荧光定量分析原理,对重组乳杆菌质粒拷贝数进行分析。借助计算机软件Oligo5对乳杆菌基因组和重组质粒DNA进行分析,设计特异性良好的引物,建立绝对定量的QPCR检测方法。利用该方法对3株重组乳杆菌质粒拷贝数进行测定。试验表明,所建立的实时荧光定量PCR检测方法能够快速、准确、特异、灵敏地确定重组乳杆菌质粒拷贝数,为进一步研究发酵过程中质粒拷贝数和表达水平相关性提供可靠基础。To establish a rapid accurate specific detecting method to analyze recombinant lactic acid bacteria expression vec-tor plasmid copy number,a real-time fluorescence quantitative PCR was develped based on the principle of SYBR Green I fluores-cent quantitative analysis.Lactic acid bacteria genomic and plasmid DNA were analyzed with the aid of computer software Oligo5.Specific primers were designed to determine recombinant lactic acid bacteria plasmid copy number by method for absolute quantifi-cation. 3 strains of recombinant lactic acid bacteria plasmid copy number were determined with the same method. The experimentshowed that established real time fluorescence quantitative PCR were able to determine recombinant lactic acid bacteria plasmidcopy number rapidly,accurately and sensitively.

关 键 词：乳杆菌 PLA 实时荧光定量PCR 

分 类 号：S852.61[农业科学—基础兽医学]