重复性
转基因水稻
生物
分子生物学
质粒
标准曲线
实时聚合酶链反应
基因
计算生物学
色谱法
遗传学
转基因作物
化学
转基因
生物化学
作者
Jaehwan Kim,Saet-Byul Park,Hyo-Jeong Roh,Min-Ki Shin,Guiim Moon,Jin-Hwan Hong,Hae‐Yeong Kim
标识
DOI:10.1016/j.foodchem.2017.01.065
摘要
One novel standard reference plasmid, namely pUC-RICE5, was constructed as a positive control and calibrator for event-specific qualitative and quantitative detection of genetically modified (GM) rice (Bt63, Kemingdao1, Kefeng6, Kefeng8, and LLRice62). pUC-RICE5 contained fragments of a rice-specific endogenous reference gene (sucrose phosphate synthase) as well as the five GM rice events. An existing qualitative PCR assay approach was modified using pUC-RICE5 to create a quantitative method with limits of detection correlating to approximately 1–10 copies of rice haploid genomes. In this quantitative PCR assay, the square regression coefficients ranged from 0.993 to 1.000. The standard deviation and relative standard deviation values for repeatability ranged from 0.02 to 0.22 and 0.10% to 0.67%, respectively. The Ministry of Food and Drug Safety (Korea) validated the method and the results suggest it could be used routinely to identify five GM rice events.
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