转人溶菌酶基因奶牛多重PCR快速检测方法的建立

建立转基因奶牛多重PCR快速检测方法,为转基因动物及产品进出境检测技术平台的建立提供技术支持,并为转基因动物及产品检测技术标准的制定提供参考。根据牛物种特异性基因线粒体DNA(mitochondrial DNA,mtDNA)设计奶牛内源性基因引物,根据外源基因人溶菌酶基因(human lysozyme,hLY)及标记基因新霉素磷酸转移酶基因(NPTⅡ)设计特异性引物,优化反应条件,建立转基因奶牛多重PCR检测方法。结果表明,建立的方法灵敏、快速、特异,一个反应可以检测多个基因片段,可有效用于转基因奶牛外源基因的检测。

Development of Multiplex PCR Detection of Human Lysozyme Gene in Transgenic Cow

For developing the typical testing standard of transgenic mammals and its derived food and the technical support for entry﹠exit animal quarantine, detection of the transgenic component in transgenic cow by multiplex PCR was developed in this study.The endogenous mitochondrial DNA(mtDNA) gene specific for cow identification, exotic human lysozyme(hLY)，marker NPTⅡ were amplified according to the differential primers. The process of the multiplex PCR reaction was optimized. The system described herein represent simple, accurate and sensitive detection methods in which only one reaction was necessary to detect multiple target sequences that could be reliably used for the identification of specific lines of transgenic cows.