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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。
In this study, we optimized the nucleic acid extraction method of different papaya materials and their processed products and successfully extracted the DNA that can be used for PCR amplification reaction. Papain specific endogenous protein PAPAIN gene fragment as an internal reference gene for testing the quality of DNA extracted effectively avoid the false negative PCR reaction and can be used to determine whether the test sample contains papaya components. The gene of coat protein (CP) and replicase (RP) of the target gene RUSDV, which is currently used widely in China, were selected as the lines for identification. This method is applicable to the inspection and quarantine of genetically modified papaya and its products by the entry-exit inspection and quarantine authorities and the agricultural authorities.