抗虫转CpTI基因成分现场可视化检测方法的建立  

Development of Visual Loop-Mediate Isothermal Amplification Assay for CpTI gene

作　　者：王永[1,2] 兰青阔 朱珠 赵新[1,3] 陈锐 刘娜 李欧静[4] 郭永泽 程奕 

机构地区：[1]天津市农业质量标准与检测技术研究所,天津300381 [2]河北农业大学植物保护学院,河北保定071001 [3]天津大学化工学院,天津300072 [4]天津农学院农学系,天津300384

出　　处：《湖南农业科学》2014年第2期22-24,共3页Hunan Agricultural Sciences

基　　金：农业科技成果转化资金项目(2011GB2A100011);科技部国际合作项目(2006DFA32380)

摘　　要：豇豆胰蛋白酶抑制剂(CpTI)基因是目前仅次于Bt基因的广谱性抗虫基因,常用于转基因水稻和转基因棉花的基因工程研究中。根据CpTI基因特异性碱基序列设计引物,应用可视化环介导等温扩增技术,对转基因水稻科丰6号进行扩增,同时建立转基因成分人工污染的食品模型,比较了LAMP法与定性PCR方法检测的敏感性。结果表明:该方法仅对CpTI基因产生特异性扩增,灵敏度达0.005%;所建立的CpTI基因现场可视化筛查方法具有较高的特异性与灵敏性,操作简单、快速,可用于含有CpTI基因转基因成分污染的快速检测。It is important to detect Cowpea Trypsin Inhibitor （CpTI） gene in food or feed materials in order to prevent the illegal diffusion. A rapid visual loop-mediated isothermal amplification （LAMP） method assay for CpTI gene detection was developed. A set of primers were designed according to the nucleotide sequence of the target CpTI gene, seven genetically modified organisms （GMO） were detected by LAMP for method specificity, and meanwhile the mode of artificially contaminated food was constructed to evaluate the sensitivity of LAMP assay and qualitative PCR method. The results showed that the CpTI gene had specific amplification, but the non- CpTI GMO submitted negative reactions. Sensitivity of LAMP assay for Kefeng No.6 genetically modified rice was 0.005%. In conclusion, the LAMP assay developed in the present study is a specific, sensitive, simple and convenient method for the rapid screening of CpTI gene in contaminated foods.

关 键 词：CPTI基因 环状等温扩增技术(LAMP) 现场可视化 

分 类 号：S339.31[农业科学—作物遗传育种]