猪IGF-Ⅰ基因荧光定量PCR标准曲线的建立  

Development of the Standard Curve for Detecting Pig IGF-Ⅰ mRNA with Fluorescence Quantitative Polymerase Chain Reaction

作　　者：肖书奇[1] 张嘉保[1] 张树敏[2] 赵志辉[3] 刘保国[4] 李爽[1] 戴立胜[1] 赵中利[1] 高妍[1] 李毅[1] 

机构地区：[1]吉林大学实验动物中心,吉林长春130062 [2]吉林省农业科学院,吉林公主岭136100 [3]吉林大学畜牧兽医学院,吉林长春130062 [4]河南科技学院动物科学学院,河南新乡453003

出　　处：《生物技术》2006年第6期52-55,共4页Biotechnology

基　　金：国家振兴东北老工业基地科技攻关项目资助(2004BA907A22)

摘　　要：目的：建立用荧光定量PER方法检测猪IGF—I基因表达量的标准曲线。方法：根据自己克隆的猪IGF—I（GenBankNo．DQTS46s7）基因mRNA序列，设计合成引物和探针，采用Taqman探针荧光定量RT—PCR的检测方法，构建检测猪IGF—I基因表达量的标准曲线。结果：由pMD-18TIGF—I所构建的标准曲线线性关系良好（反应体系中含1×102～1×10＇拷贝猪IGF—I基因分子时，扩增反应ct值与拷贝数的对数呈线性关系）、灵敏度高（可检测出低于10个拷贝，肚的样品）、特异性强、准确可靠。结论：成功构建了用荧光定量RT—PCR方法检测猪IGF—I基因表达量的标准曲线。Objective： The standard curve for detecting pig IGF- Ⅰ mRNA expression with fluorescence quantitative polymerase chain reaction was developed.Methods：Tile probe and primers were designed and synthesized according to the pig IGF - Ⅰ （GenBank No. DQ784687）mRNA sequence. Then a Taqman probe fluorescence quantitative RT - PCR assay was carried, and the standard curve for detecting pig IGF - Ⅰ mRNA expression was constructed.Results：The standard curve made by pMD- 18T IGF- Ⅰ had good linear dependence,and it was sensitive（it could detect 〈 10 copies/μL of plasmid DNA） and specific. Conclusion： The standard curve for detecting pig IGF - Ⅰ mRNA expression with fluorescence quantitative RT- PCR was developed successfully.

关 键 词：猪 胰岛素样生长因子-I TAQMAN探针 荧光定量RT—PCR 标准曲线 

分 类 号：Q781[生物学—分子生物学]