摘要
目的:建立转Bt基因稻米的降落PCR-毛细管电泳快速定性检测方法。方法:转Bt基因稻米经磨碎后提取基因组DNA,对其内源蔗糖磷酸合酶SPS基因和抗虫毒蛋白Bt基因同时进行聚合酶链反应(PCR)扩增,产物用无胶筛分毛细管电泳分离、激光诱导荧光检测器检测。结果:在优化条件下,25 min内即可完成SPS和Bt基因的检测,日内精密度在2.37%～3.19%之间,特异性实验结果表明本法特异性较高。结论:所建立的方法快速、分离效能高、样品用量少且成本低廉,适用于转Bt基因稻米的快速鉴定。
Objective： To establish a rapid,sensitive and reliable method for detection of genetically modified rice with Bt gene.Methods： The touchdown polymerase chain reaction（PCR） with two primer pairs targeted to the endogenous control gene（SPS） and cultivar-specific gene（Bt） was performed after DNA was extracted.The PCR products then were separated by non-gel sieving capillary electrophoresis and detected by laser-induced fluorescence detector.Results： The detection of the PCR products was completed within 25 minutes under the optimized conditions.The within-day precision of Migration time was 2.37%~3.19%.The results of specificity tests demonstrated that the method was highly specific.Conclusion： Comparing to traditional gels electrophoresis,the proposed method requires smaller amount of samples,which is more rapid,sensitive and accurate.Thus it offers an efficient alternative method to routinely monitor genetically modified rice with Bt gene.
出处
《中国卫生检验杂志》
CAS
北大核心
2012年第11期2531-2533,共3页
Chinese Journal of Health Laboratory Technology
基金
国家自然科学基金(青年基金)项目(81102162)
四川大学青年教师启动基金项目(2010SCU11013)
关键词
转Bt基因稻米
降落PCR
毛细管电泳
激光诱导荧光检测
Genetically modified rice with Bt gene
Touchdown polymerase chain reaction（PCR）
Capillary electrophoresis（CE）
Laser induced fluorescence detection
