作　　者： ; ; ;

机构地区： 华南农业大学资源环境学院

出　　处： 《湖南农业大学学报（自然科学版）》 2010年第6期626-629,682,共5页

摘　　要： 为监测和管理转基因番木瓜华农1号的商品化生产、销售,建立了相应的转基因定性检测方法.选用木瓜蛋白酶作为番木瓜的内源对照基因,建立对转基因番木瓜PRSV-Rep基因、外源CaMV35S启动子序列、NOS终止子序列和NPTⅡ基因的常规PCR检测体系.退火温度为40～60℃时,都分别扩增出了清晰的目的基因.单基因常规PCR检测转基因模板量的下限为5×10-7g.建立的三重PCR检测技术,检测了包括NPTⅡ、Rep和CaMV35S;NPTⅡ、Rep和Nos;NPTⅡ、Rep和Papain等3个组合.建立了NPTⅡ、ReP、CaMV35S和Nos4个基因的四重PCR检测技术. A set of transgenic detection methods and technology had been established to supervise and manage the commercial producing and selling of the transgenic papaya-Huanong No.1 Papain（Pn）was chosen as the endogenous comparison genes of papaya and primers were designed to detect the transgenic papaya and non-transgenic papaya.The regular PCR was established to detect the PRSV-Rep,exogenous CaMV 35S promoter sequence,NOS terminator sequence and the NPTⅡ of transgenic papaya.The clear bands were amplified when the annealing temperature was 40 ℃ to 60 ℃.The sensitivity of the single gene was 5×10-7 g.The Triple-gene-PCR detection system was created to detect three combination genes of NPTⅡ,Rep and CaMV 35S;or NPTⅡ,Rep and Nos;or NPTⅡ,Rep and Papain.And the Quad-PCR system was built to test four genes of NPTⅡ,Rep,CaMV35S and Nos as well.

关 键 词： 番木瓜环斑病毒 转基因番木瓜 多重聚合酶链式反应

领　　域： [生物学]