转人血清白蛋白基因奶牛多重PCR快速检测方法的建立

Multiplex PCR Detection of Exogenous Gene in α-LA Transgenic Cow

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监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。根据转基因奶牛内源基因β2-微球蛋白基因(β2-microglobulin,B2 M),外源基因人血清白蛋白基因(Human α-lactalbumin,α-LA)及标记基因绿色荧光蛋白基因(EGFP)和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。该方法敏感、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。

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 B2M gene,exoticα-LA,marker EGFP、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 is necessary to detect multiple target sequences that can be reliably used for the identification of specific lines of transgenic cows.