pIRES2-EGFP-MyoG真核表达载体的构建和鉴定  

Construction and identification of eukaryotic expression vector pIRES2-EGFP-MyoG

作　　者：徐婷婷[1] 李树峰[1] 佟慧丽[1] 冯霞[1] 卢丽[1] 严云勤[1] 

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030

出　　处：《黑龙江畜牧兽医》2010年第10期15-18,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：转基因生物新品种培育科技重大专项(2008ZX08007-002)

摘　　要：为了构建能够表达日本和牛生肌素(myogenin,MyoG)的真核细胞表达载体pIRES2-EGFP-MyoG,试验从日本和牛成纤维细胞基因组中扩增出MyoG基因,插到带有EGFP和内部核糖体转入位点(IRES)的真核细胞表达载体pIRES2-EGFP中,经酶切鉴定正确后转染鲁西黄牛胎儿成纤维细胞,用RT-PCR和Western-blot检测MyoG基因的表达情况。结果表明:真核表达载体pIRES-EGFP-MyoG构建成功,且在鲁西黄牛胎儿成纤维细胞中得到了有效表达。说明真核表达载体中的MyoG能够在鲁西黄牛胎儿成纤维细胞中成功表达。To construct the eukaryotic expression vector of wagyu Myogenin（myogenin,MyoG） gene,the experiments uses wagyu fibroblasts genomic DNA to amplify MyoG gene.MyoG gene was inserted into the eukaryotic expression vector pIRES2-EGFP with EGFP and the internal ribosome site（IRES）,is was transfected into fetal fibroblast cells of Luxi huangniu identification with double restriction enzyme digestion,MyoG gene expression were detected by RT-PCR and Western blot.The result showed that the constructed eukaryotic expression vector pIRES-EGFP-MyoG wer confirmed by restriction enzymolysis and could expressed efficiently in fetal fibroblast cells of Luxi huangniu,MyoG was expressed in them successfully.It may lay a good foundation to further study MyoG for biologzcal function and mechanism.

关 键 词：日本和牛 MYOG 转染 PIRES2-EGFP 

分 类 号：S823.92[农业科学—畜牧学]