双引物PCR检测抗草甘膦豆粕饲料中的转基因成分.pdfVIP

, 2008, 22( 6): 17~ 19 Journal of Anhu i Science and T echnology Un i ersity PCR 1, 3 2 3 朱元招 , 尹靖东, 尚秀国 ( 1. , 233100; 2. , 100094; 3. , 100085) : 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 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 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 : 豆粕; 双引物 PCR; 抗草甘 大豆; 转基因成分; 饲料 : S816. 17; S816. 35 : A : 1673- 8772( 2008) 06- 0017- 03 D etection of Transgenic Component in G lyphosate- tolerant Soybean M eal w ith Double- prim er PCR 1, 3 2 3 ZHU Yuan- zhao , YIN Jing- dong , SHANG X iu- guo ( 1. An mi al Science College, Anhui Science and Technology U ni ersity, Fengyang 233100, China; 2. Ch inaA griculturalUn i ersity, Beijing 100094, Ch ina; 3. Beijing GendoneAgriculture Science and Technology Co. Ltd. , Beijing 100085, China) A bstract: Soybean lec gene as internal reference and CaMV35s promoter, nos term inator or cp4 epsps gene in exogenous gene cassette inserted in glyphosate- tolerant soybeansw ere synchronously amplified in one PCR reac- tion to de elop a double- prmi er PCR m ethod for detection of transgen ic soybeans. PCR productsw ere cutw ith restr icti e endonuclease. The results show ed that lec gene and the specific exotic gene w ere amp lified in the standard transgene- positi e soybeans, A rgentina soybeans, RR soybeans, RR soybean mealw ith double- prmi - er PCR. Further, the fragments were confirmed to be the target genes w ith the same length as expected by