实时荧光定量PCR法检测抗虫棉历基因拷贝数方法的建立.pdfVIP

ofAnhui 安徽农业科学。Journal Agri．Sci．2008，36【26)：11376—11377 责任编辑张彩丽责任校对傅真治 实时荧光定量PCR法检测抗虫棉历基因拷贝数方法的建立 闫喜中1”，张锐1，孟志刚1，孙国清1i周涛1，郭三堆1* 摘要 632X+43．783 基因拷贝数与D值的关系为Ct=一3．549 512，相关系数∥为0．997 867。【结论]实时荧光定量王)CR检测转基因抗虫棉 鼠基因的方法具有特异性好、灵敏度高的特点。 关键词助基因；实时荧光定量PCR；标准曲线 中图分类号$562文献标识码A 文章编号0517—661l(2008)26—11376—02 aReal-time PCR for the NumberofBtGenein Insect-resistantCotton Establishing QuantitafiveAssayDeletingCopy Transgenic YAN et Research of for GeneRe— Institute，ChineseAgricultureSciences，NationalKeyFacilityCrop Xi-zhongal(Biotechnology Academy and l 0008 sourceGenetic 1) Improvement．Beijing aimofthis wasto realaime PCR for Bt in insect-resist— Abstract0bjectivene paper develop quantitativeassaydetectinggenetransgenic 111e ofBt thesoftware 5．0．Thestandardcurvewas based antcotton．『Mcthod and were primersprobes gene designedusing primer prepared ForBt 632X onthelinear betweentheamountof Result relationship inputplasmidDNA(x)andcyclethreshold(Ct)．I gene，Ct=一3．549 an 十43．783 correlationCOCmcientR。was0．997 ConclusionThereal-time PCRmethodcouldbeusedas effective 512，and 867．f quantitive detectionand methodforBt anditisamenableto forits and quantification gene high-throughoutassay specificity，sensitivityrapidity． wordsBt curve Key gene：Real-timequantitativePCR；Standard Southern印迹方法是最常用的转基因植株外源基冈拷贝