微生物酶制剂中转基因微生物的实时荧光PCR检测方法

A method of real-time PCR for detection of genetically modified microorganisms in microbiology derived enzyme

针对微生物酶制剂中残留转基因微生物的检测问题,根据醇氧化酶-1(AOX1)启动子基因的序列信息设计一对引物及一条MGB探针,建立了转基因微生物的实时荧光PCR筛选检测方法。实验结果表明,该方法灵敏度达到1pg,可以准确判定生产线上不同分离阶段的微生物酶制剂中的转基因微生物残留情况。该方法准确度和灵敏度高,操作方便,可作为微生物酶制剂中转基因微生物分离状况的监测方法,也可为其他转基因微生物检测研究提供借鉴和参考。

A new method of real-time PCR was built to detect genetically modified microoganisms in microbioloogy derived enzyme. The primer and the MGB probe were desgined on the sequence of the alcohol oxidase-1 （AOX1） promoter. The experimental result showed that the sensitivity of detction was lpg Pichia pastoris DNA, and could accurately analyze the residue status of genetically modified microorganism in microbiology derived enzyme from different separation stage. This practical method could be used to monitor separation of transgenic mocrobiology in microbiology dereirved enzyme, and provide reference for the detction of genetically modified microorganism related stdudies.