双引物PCR检测抗草甘膦豆粕饲料中的转基因成分

Detection of Transgenic Component in Glyphosate-tolerant Soybean Meal with Double-primer PCR

在一个PCR反应体系内,同步扩增大豆内参照基因lec和抗草甘膦大豆外源基因盒中的CaMV35s启动子、nos终止子或cp4 epsps基因,建立转基因大豆双引物PCR检测方法。PCR产物用限制性内切酶酶切进一步鉴定。结果表明,用双引物PCR扩增出标准转基因大豆阳性对照、阿根廷大豆、美国RR大豆及豆粕中的lec基因和相应的特异性外源基因;酶切反应确证PCR产物为特定片段大小的目的序列。建立的双引物PCR法适用于抗草甘膦大豆原料和豆粕饲料中转基因成分的简单化、高精度和高灵敏检测。

Soybean lec gene as internal reference and CaMV35s promoter, nos terminator or cp4 epsps gene in exogenous gene cassette inserted in glyphosate - tolerant soybeans were synchronously amplified in one PCR reaction to develop a double - primer PCR method for detection of transgenic soybeans. PCR products were cut with restrictive endonuclease. The results showed that lec gene and the specific exotic gene were amplified in the standard transgene - positive soybeans, Argentina soybeans, RR soybeans, RR soybean meal with double - primer PCR. Further, the fragments were confirmed to be the target genes with the same length as expected by enzyme incision reaction. Therefore, the developed double - primer PCR is a simple, high - precision and sensitive method for detecting transgenic component in glyphosate -tolerant soybean and soybean meal.