番木瓜及其深加工制品中转基因成分定性PCR检测方法的建立(英文)

本研究对番木瓜不同材料及其深加工制品的核酸提取方法进行了优化,成功提取出可用于PCR扩增反应的DNA。选取番木瓜特异性的内源蛋白PAPAIN基因片段作为内参基因,用于检验提取DNA的质量,有效避免了PCR反应的假阴性并能用于确定检测样品中是否含有番木瓜成分。选取国内目前应用比较多的外源目的基因环斑病毒外壳蛋白(CP)基因以及复制酶(RP)基因作为品系鉴定的基因。本方法适用于出入境检验检疫部门和农业部门对转基因番木瓜及其制品的检验监测。

Development Study on PCR Detection Method for Genetically Modified Components in Papaya and Its Highly-processed Products

This research has optimized the method of extracting the DNA of different parts of papaya and its highly-processed products, therefore successfully extracted the DNA for PCR(polymerase chain reation). Papain gene,a papaya species-specific protein gene is adopted as an internal control to access the quality of the DNA extracted so as to avoid the pseudo-negative case of PCR reaction and to confirm the existence of papaya in the samples as well. In this research, the genes applied are PRSV (papaya-ring-spot-virus) CP(coat protein) gene and RP(replicase) gene. This method is applicable and reliable for the determination and supervision of GM papaya and its processed products for Bureau of Entry-Exit Inspection and Bureau of Agriculture.