作物学报2018，Vol.44Issue(7)：949-955,7.DOI:10.3724/SP.J.1006.2018.00949
抗病转基因水稻M12及其产品成分的定性、定量PCR检测方法
A Qualitative and Quantitative PCR Detection Method for Disease-resistant Genetically Modified Rice M12 and Its Derivates
摘要
Abstract
In this study, the specific sequence of genetically modified Rice M12 and the endogenous reference gene sps were amp-lified to construct a T vector as the plasmid pM12 for establishing the qualitative and quantitative PCR detection method of trans-genic Rice M12 and its derivates. The qualitative PCR method could specifically quantify the samples of M12 with the detection sensitivity about 100 copies of the rice haploid genome. On the basis of SYBR Green qPCR assay, R2 values of standard curves of M12 and sps were 0.998 and 0.997, the amplification efficiency was 95.3% and 108.4%, respectively. Moreover, the standard deviations (SD) of repeatability ranged from 0.043 to 0.276. The limit of quantification (LOQ) and limit of detection (LOD) were 100 and 10 copies, respectively. The mixed rice sample containing 1.0% gene transforming into rice was exactly quantified by the developed quantitative PCR method, and the quantified bias between the true value and tested value was below 8.0%. In conclu-sion, these methods can be used for identifying and quantifying M12 and its derivatives.关键词
品系特异性/定性PCR/定量PCR/M12Key words
event-specific/qualitative PCR/quantitative PCR/M12引用本文复制引用
李鹏,张琳,叶吉妮,贺诗瑶,贾军伟,潘爱虎,唐雪明..抗病转基因水稻M12及其产品成分的定性、定量PCR检测方法[J].作物学报,2018,44(7):949-955,7.基金项目
本研究由国家自然科学基金项目(31500461), 上海市科技兴农重点攻关项目[沪农科攻字(2015)第 4-3], 上海市农业科学院卓越团队项目[农科创2017(B-07)]和上海市农业转基因生物安全监管专项资助.This study was supported by the National Natural Science Foundation of China (31500461), the Key Technologies Program of Shanghai Agricultural Commission [2015(4-3)], SAAS Program for Excellent Research Team [2017(B-07)], and the Agricultural GMO Safety Supervi-sion Program of Shanghai. （31500461）
