植物油DNA的高效提取方法研究  

High efficient extraction of DNA from plant oil

作　　者：蔡翠霞[1] 肖维威[1] 吴慧慧[1] 周琳华[1] 吴永彬[1] 耿启斌 马文丽[1] 

机构地区：[1]南方医科大学基因工程研究所,广州510515

出　　处：《中国油脂》2014年第1期69-72,共4页China Oils and Fats

基　　金：国家公益性行业科研专项(200910265);广东省科技计划项目(2008B080701027)

摘　　要：针对植物油中DNA含量极低、DNA序列片段短、破坏严重的特点,以食用植物油为原料,旨在建立一种从植物油中稳定、高效提取DNA的方法。实验采用硅膜吸附柱法提取植物油基因组DNA,PCR扩增其相应内源基因进行质量鉴定,再针对通用的外源基因CaMV35S启动子进行PCR、LAMP和实时荧光PCR检测对该方法进行评价。结果表明:该方法提取的DNA质量可靠,采用PCR、LAMP和实时荧光PCR技术成功地检测出了植物油中的CaMV35S启动子。因此,硅膜吸附柱法提取的植物油DNA可以作为PCR、LAMP、实时荧光PCR扩增模板用于转基因成分的检测,该方法经济、稳定、安全,为植物油的基因检测奠定了基础。According to the features of low content, short sequence fragment and serious destruction of DNA in plant oil, a stable and high efficient extraction method of DNA from edible plant oil was estab lished. DNA was extracted from plant oils using silicon film adsorption column method, and the endoge nous genes in the DNA were amplificated by PCR to identify DNA quality, then the exogenous gene pro moter CaMV355 was detected by PCR, LAMP and real -time fluorescent PCR to evaluate the method. The results showed that the DNA extracted by the method had reliable quality and it was successful to de tect the promoter CaMV355 by PCR, LAMP and real -time fluorescent PCR. The DNA extracted from plant oil by the method could be used as the amplification template of PCR, LAMP and real - time fluo rescent PCR for genetically modified ingredients detection. The method had items of stableness, effective- ness and safety which could provide the foundation for plant oil gene detection.

关 键 词：植物油 DNA提取 转基因检测 

分 类 号：TS225.1[轻工技术与工程—粮食、油脂及植物蛋白工程]