转基因玉米筛选检测阳性标准分子的构建和应用

Construction and Application of Positive Standard Molecular for Screening Detection of Genetically Modified Maize

为了满足转基因玉米成分检测中阳性对照不易获得及不易长久保存的缺陷,通过分析公共数据库资源,建立了包含玉米内标准基因z SSIIb及遗传转化中常用的Ca MV35S启动子、nos启动子、Ca MV35S终止子、nos终止子的的标准分子。灵敏度检测发现,在检测体系中含有1 000个分子时,可以非常清楚扩增到所有5个片段的目的条带。随机挑选我国批准进口的7种转基因玉米,对标准分子进行验证,结果表明,在相应的阳性样品和标准分子中都能扩增到特异的目的条带,构建的标准分子可以满足作为转基因玉米筛选检测中阳性对照的要求。

In order to solve lack of the positive control, a positive standard molecular which containing maize endogenous gene zSSIIb, CaMV35S promoter, nos promoter, CaMV35S terminator, and nos terminator was construct- ed. Sensitivity detection of the standard molecular indicated the electrophoretic lines were very clearly when the number of the template was above 1 000 molecular. Specificity testing demonstrated that, right bands in the stan- dard molecular and the positive samples could be obtained. The above results revealed the effectiveness of the stan- dard molecular for GMO screening.