Long Likun, Xing Zhenjuan, He Yuxuan, Yan Wei, Li Congcong, Xia Wei, Dong Liming, Zhao Ning, Ma Yue, Xie Yanbo, Liu Na, Li Feiwu
Institute of Agricultural Quality Standard and Testing Technology, Jilin Academy of Agricultural Sciences, Changchun 130033, China.
Foods. 2022 Aug 22;11(16):2535. doi: 10.3390/foods11162535.
As an effective tool for genetically modified organism (GMO) quantification in complex matrices, digital PCR (dPCR) has been widely used for the quantification of genetically modified (GM) canola events; however, little is known about the quantification of GM canola events using endogenous reference gene () characteristics by dPCR. To calculate and quantify the content of GM canola using endogenous reference gene () characteristics, the suitability of several ERGs of canola, such as cruciferin A (), acetyl-CoA carboxylase (), phosphoenolpyruvate carboxylase (), cruciferin storage (), oleoyl hydrolase (), and high-mobility-group protein I/Y (), was investigated by droplet dPCR. and were more specific and stable in copy number in the genome of L. than the other genes. By performing intra-laboratory validation of the suitability of ERG characteristics for the quantification of GM canola events, the ddPCR methods for and were comprehensively demonstrated in dPCR assays. The methods could provide technical support for GM labeling regulations.
作为在复杂基质中对转基因生物(GMO)进行定量的有效工具,数字PCR(dPCR)已被广泛用于转基因油菜事件的定量;然而,关于利用内源参考基因()特征通过dPCR对转基因油菜事件进行定量的研究却很少。为了利用内源参考基因()特征计算和定量转基因油菜的含量,通过液滴dPCR研究了几种油菜内源参考基因的适用性,如十字花科蛋白A()、乙酰辅酶A羧化酶()、磷酸烯醇式丙酮酸羧化酶()、十字花科蛋白贮藏基因()、油酰水解酶()和高迁移率族蛋白I/Y()。与其他基因相比,和在甘蓝型油菜基因组中的拷贝数更具特异性和稳定性。通过对用于转基因油菜事件定量的内源参考基因特征适用性进行实验室内部验证,在dPCR分析中全面展示了针对和的液滴数字PCR方法。这些方法可为转基因标识法规提供技术支持。