SYBR GreenⅠ荧光定量PCR法检测转双基因猪pGH基因与IGF-1基因的表达  

The PGH Gene and IGF-1 Gene Expression Detected by SYBR Green I Real-time Quantitative PCR in Double Transgenic Pigs

作　　者：胡义洁 吴明明[2] 朱艳菲[1] 孙金海[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]中国农业大学,北京100193

出　　处：《中国畜牧杂志》2014年第13期16-21,共6页Chinese Journal of Animal Science

基　　金：转基因生物新品种培育重大专项课题研究计划(2013ZX08006-003);山东省现代农业产业技术体系生猪创新团队建设项目

摘　　要：为确定猪生长激素基因(pGH)与胰岛素样生长因子-1(IGF-1)在F0代保育期转双基因猪和同窝非转基因猪体内mRNA水平的表达差异情况,利用SYBR GreenⅠReal-time PCR法检测精子介导纳米基因载体法获得的F0代2月龄转基因猪pGH基因与IGF-1基因的mRNA表达水平,根据荧光定量结果绘制熔解曲线和扩增曲线图谱,分析图谱并用F=(1+E)-ΔΔCt公式计算其相对表达量。结果表明:F0代转双基因猪pGH基因的mRNA表达量是同期非转基因猪的1.2倍,IGF-1基因的mRNA表达量是同期非转基因猪的1.53倍,说明这2种外源基因均成功导入,并能在猪体内表达。本实验建立了能够稳定测评家猪生长激素与胰岛素样生长因子-1基因表达水平的方法,为进一步分析pGH-IGF-1生长轴对家猪生产性能的具体影响及转基因的实际应用提供了分子生物学方面的资料。In order to determine the difference of mRNA expression level in exogenous porcine growth hormone gene （pGH） and insulin-like growth factor-1 （IGF-I） between F0 generation of double transgenie pigs and brood non-transgenic pigs of nursery phase, pGH gene and IGF-1 gene in transgenic pigs of 2-month-old were detected by SYBR Green [ Real-time Fluorescence Quantitative PCR method, which were made by nano-gene carrier method induced by sperm and normal boar. According to amplification curve and melting curve maps, relative transcript level was gained through the equation F=（I＋E）^-△△Ct. The result indicated that the mRNA expression level of pGH gene in F0 generation of double transgenic pigs was 1.2 times as non-transgenic pigs, and that of IGF-1 gene was 1.53 times as non-transgenic pigs. The results showed that the foreign pGH gene and IGF-1 gene were expressed successfully in pigs. A set of molecule biology method of evaluating pGH and IGF-1 relative expression level stably was built, and provided molecular biology information for further analysis of the specific influence of pGH-IGF-1 axis on pig production performance and the practical application of genetically modified.

关 键 词：转双基因 猪生长激素基因 胰岛素样生长因子-1基因 实时荧光定量PCR 

分 类 号：S828.2[农业科学—畜牧学]