PCR结合核酸斑点杂交技术检测转基因番茄研究  

Research on Detection of Transgenic Tomato with PCR in Combination with Nucleic Acid Spot Hybridization Technology

作　　者：梁彦君[1] 陈文虎[1] 王梦娜[1] 许爱霞[1] 冯俊丽[1] 

机构地区：[1]浙江理工大学生命科学学院生物工程研究所,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2013年第5期747-752,共6页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家质检总局科研项目(2012IK290)

摘　　要：基于转基因番茄的安全性评价,建立健全番茄转基因检测体系尤为迫切。文章采用转基因作物检测常用的传统定性PCR技术和核酸斑点杂交技术,针对转基因番茄中常用的35S(花椰菜花叶病毒启动子)、NOS(胭脂碱合成酶终止子),NPTII(新霉素转移酶基因),35S-EFE(35S和番茄EFE基因的交联结构)、以及Lat52(番茄内参基因),检测了阳性对照质粒PBI121、华蕃一号样品和阴性对照合作903番茄。从华蕃一号中成功检出35S、NOS、NPTII、35S-EFE和内参基因Lat52,阴性对照番茄合作903中只有内参基因Lat52检出。试验重复性良好,检测灵敏度高,两种方法相互补充,排除假阳性。核酸斑点杂交一次可检出多个外源基因,增加了检测通量,为我国番茄转基因检测提供参考。It is especially urgent to establish a complete tomato transgenic detection system based on transgenic tomato safety evaluation. This paper detects positive control plasmid PBI121, No. 1 Huafan sample and negative control cooperation 903 tomato with the traditional PCR technology and nucleic acid spot hybridization technology commonly used in detection of transgenic crops in allusion to 35S（cauliflower mosaic virus promoter）, NOS（nopaline synthase terminator）, NPTII（neomycin transferase gene）, 35S- EFE（35S and cross-linked structure of tomato FEE gene） and Lat52（tomato reference gene） commonly used in transgenic tomato. 35S, NOS, NPTII, 35S-EFE and reference gene Lat52 are detected in No. 1 Huafan and only reference gene Lat52 is detected in negative control tomato cooperation 903. The test has a good repeatability and high detection sensitivity. Both methods complement each other and eliminate false positive. Nucleic acid spot hybridization can detect multiple foreign genes once and increases the detection flux and provides reference for tomato transgenic detection in China.

关 键 词：转基因番茄 外源基因检测 定性PCR技术 核酸斑点杂交 

分 类 号：Q788[生物学—分子生物学]