转基因番茄中外源目的基因检测方法的筛选  

Screening of detection method of the exogenous gene from vaccine of the Transgenic tomato

作　　者：白国辉[1] 田源[2] 白朋元[2] 韩琪[2] 刘建国[2] 

机构地区：[1]遵义医学院医学与生物学研究中心,贵州遵义563000 [2]遵义医学院口腔学院口腔内科学教研室,贵州遵义563000

出　　处：《遵义医学院学报》2011年第5期457-460,共4页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:30160086);贵州省优秀科技人才省长专项基金项目(NO:黔省专合字[2006]44);贵州省科技攻关项目(NO:黔科合NY字[2006]3043)

摘　　要：目的用普通PCR和real-time PCR两种方法检测转基因番茄中的外源基因,通过优化条件建立一种灵敏、准确检测转基因番茄中外源基因的方法。方法以防龋用转基因番茄为实验材料,通过SDS和植物基因组DNA提取试剂盒两种方法提取植物总DNA,用普通PCR和real-time PCR检测转基因番茄中的外源目的基因。结果植物基因组DNA提取试剂盒提取样品在琼脂糖凝胶电泳图上出现单一条带,条带清楚无拖尾,而用SDS法检测时常会出现拖尾现象。用普通PCR法重复检测40株转基因番茄平均检出率为88.75%,出现假阴性结果,而用real-timePCR法检测相同样本未出现假阳性或假阴性结果,检出率为100%。结论植物基因组DNA提取试剂盒结合real-time PCR技术是检测转基因番茄中外源目的基因的一种准确、灵敏的方法。Objective To establish a sensitive and accurately method that detect the exogenous gene from transgenic tomato after the exogenous gene of transgenic tomato were detected and optimized by ordinary PCR and real-time PCR,Methods Extracted the total plant genom of the transgenic anti-caries tomato by SDS and the kit,detected the exogenous gene of transgenic tomato by ordinary PCR and real-time PCR.Results There was a single clear band without tail appeared on the agarose gel if the total plant genom of the transgenic anti-caries tomato was extracted by Genomic DNA extraction kit.While,the bands were often with tail by the SDS method.It was often occured false negative or false positive amplification if the exogenous gene of transgenic tomato was repeatedly detected by ordinary PCR,but it never occur by real-time PCR Conclusion Extraction of the total plant genom of the plant by kit and real-time PCR technology is a sensitive and accurate method to detect the exogenous gene from the transgenic anti-caries tomato.

关 键 词：转基因番茄 防龋疫苗 REAL-TIME PCR PCR 

分 类 号：R349.83[医药卫生—基础医学]