中国组织工程研究Issue(37)：6645-6651,7.DOI:10.3969/j.issn.2095-4344.2013.37.017
Myod1和Myog真核共表达载体构建及鉴定
Construction and identification of eukaryotic co-expression vector carrying Myod1 and Myog
摘要
Abstract
BACKGROUND:Now it has cooperation and facilitative effete between myogenic regulatory factors through a long time study. So, gene therapy of double genes of Myod1 and Myog can obtain better effect, and can provide a new way for preventing denervated skeletal muscle atrophy. <br> OBJECTIVE:To construct eukaryotic co-expression vector carrying Myod1 and Myog genes. <br> METHODS:Ful-length Myod1 gene and Myog gene cDNA were amplified by reverse transcription PCR, and then inserted into pVAX1 vector after digested to establish the recombined Myod1 and Myog eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and then identified with gene sequencing. The in vitro cultured 3T3 cel s were transfected with pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and the expressions of Myod1 and Myog genes in the 3T3 cel s were detected with western blot assay in order to identify whether the 3T3 cel s could express the target protein correctly. <br> RESULTS AND CONCLUSION:The sequencing results showed that the sequence length and base sequence of Myod1 and Myog cDNA in eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP were identical with the reported sequences. Myod1 and Myog protein band expressions were detected in 3T3 cel s by western blot after transient transfection. The pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, a eukaryotic co-expression vector of Myod1 gene and Myog gene is successful y constructed.关键词
组织构建/组织构建基础实验/Myod1/Myog/真核共表达载体/基因转染/基因测序/失神经支配/骨骼肌萎缩/国家自然科学基金分类
医药卫生引用本文复制引用
高宏飞,梁炳生,双卫兵..Myod1和Myog真核共表达载体构建及鉴定[J].中国组织工程研究,2013,(37):6645-6651,7.基金项目
国家自然科学基金面上项目(81000805)@@@@General Project of National Natural Science Foundation of China, No.81000805 （81000805）
