转基因水稻标准质粒分子的构建与分析

发布时间：2018-08-08 12:59

【摘要】：针对目前转基因(GM)产品检测标准分子的缺乏,构建适用于转基因水稻的标准质粒分子pMDSPS和pMDTheli,其中包含水稻内源基因SPS通用序列,耐盐基因ThST3和Ubiquitin启动子序列的构建特异性Theli序列。对其特异性及制备标准曲线的可靠性进行鉴定研究,研究结果显示,在进行普通PCR扩增时,采用质粒标准分子为阳性对照,均能特异扩增目的条带。以标准质粒分子构建的标准曲线,SPS基因的定量标准曲线的相关系数平均为0.9993,PCR扩增效率在1.0004~1.034之间;ThST3基因的定量标准曲线的相关系数平均为0.9937,PCR扩增效率在0.9358~0.9486之间。因此,构建的标准分子pMDSPS和pMDTheli既能作为普通PCR的阳性对照,也能用于定量标准曲线的构建,为转基因水稻的定量检测提供简便、价格低廉的标准分子打下基础。
[Abstract]:In view of the lack of standard molecules for the detection of transgenic (GM) products, the standard plasmid molecules pMDSPS and pMDTheli, which are suitable for transgenic rice, are constructed, which contain the common SPS sequence of rice endogenous gene. Construction of specific Theli sequence of salt tolerant gene ThST3 and Ubiquitin promoter. The specificity and reliability of the preparation standard curve were studied. The results showed that the target bands could be amplified by using plasmid standard molecule as positive control. The correlation coefficient of the quantitative standard curve of SPS gene constructed from standard plasmid molecule was 0.9993H PCR amplification efficiency between 1.00041.034 and 0.9937%. The average PCR amplification efficiency of ThST3 gene was between 0.9358and 0.9486. Therefore, the constructed standard molecule pMDSPS and pMDTheli can be used not only as positive control of ordinary PCR, but also in the construction of quantitative standard curve, which provides a simple and inexpensive standard molecule for quantitative detection of transgenic rice.
【作者单位】：农业部农作物种子质量监督检验测试中心(深圳)/深圳市农业科技促进中心;
【基金】：深圳市技术创新项目(CXZZ20120614165508810) 转基因新品种培育重大专项(2009ZX08001-023B)
【分类号】：S511

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