稻谷中转Bar基因水稻含量的实时荧光定量检测  

Quantitative Detection of Transgenic Bar Gene Rice Grain Content in Mixed Rice Grain

作　　者：林波[1] 谭支良[1] 肖国樱[1] 汤少勋[1] 孙志洪[1] 王敏[1] 

机构地区：[1]中国科学院亚热带农业生态研究所农业生态工程重点实验室

出　　处：《农业现代化研究》2008年第5期618-621,共4页Research of Agricultural Modernization

基　　金：中国科学院知识创新工程领域前沿项目(0751012010);中国科学院知识创新工程重要方向项目(KSCX2-YW-N-49)

摘　　要：本研究以转Bar基因稻谷外源基因片断(Bar,CaMV35s)为检测对象,应用实时荧光定量PCR(RT-PCR)法建立稻谷中转Bar基因稻谷成分的定量检测方法。分别设计两对内参基因(SPS,RBE4)引物和外源基因(Bar,CaMV35s)引物,应用RT-PCR法对混合稻谷中不同量转Bar基因稻谷(30%、20%、10%、5%、0.9%、0.5%)进行定量检测,检测结果分别为:1、SPS与Bar基因组34.22%、22.14%、11.36%、6.21%、0.74%、0.32%;2、RBE4与Bar基因组28.68%、17.26%、11.92%、6.24%、0.75%、0.35%;3、SPS与CaMV35s基因组32.23%、21.87%、11.15%、4.35%、0.7%、0.37%;4、RBE4与CaMV35s基因组32.35%、22.54%、8.62%、4.62%、0.71%、0.36%。综合分析得出SPS与CaMV35s基因组合对转Bar基因稻谷定量检测的结果较为准确,可见RT-PCR法可以作为定量检测稻谷中转Bar基因稻谷成分的可信方法之一,但其准确性随转Bar基因成分含量的降低而降低,且与所选择的内参基因和外源基因引物有关。This study explored the quantitative detection of transgenic Bar gene rice grain content by detect Bar gene and CaMV35s gene fragments using RT-PCR method, we designed four couples primers for transgenic Bar gene rice content detect in rice grain by using RT-PCR. The result showed that Quantitative detection of different transgenic Bar gene rice grain content （30%. 20%, 10%. 5%. 0.9%. 0.5%） using different reference and foreign gene combination is SPS/Bar34.22%. 22.14%. 11.36%.6.21%. 0.74%, 0.32% ： RBE4/Bar 28.68%. 17.26%. 11.92%. 6.24%. 0.75%. 0.35% ; SPS/CaMV35s 32.23%. 21.87%. 11.15%. 4.35%, 0.79%. 0.37% ： RBEA/CaMV35s 32.35%. 22.54%. 8. 62%. 4.62%. 0.71%. 0.36% respectively. It can be seen that SPS/CaMV35s gene combination has smaller quantitative detection error compare with other three gene combinations. This study indicate that RT-PCR can be a reliable method for quantitative detect transgenic Bar gene rice content in rice grain, but the accuracy is relate with gene primer and decrease with the content oftransgenic content in rice grain.

关 键 词：转基因水稻 BAR基因 定量检测 实时荧光定量PCR 

分 类 号：S511[农业科学—作物学]