CMV与SP双启动子增强外源基因在小鼠骨骼肌中的表达效率  

Two Promoters CMV and SP Enhance the Expression Efficacy of an Exogenous Gene in Skeletal Muscle of Mice

作　　者：章倩倩[1] 刘松财[1] 戴建威[1] 任晓慧[1] 张永亮[1] 

机构地区：[1]吉林大学农学部畜牧兽医学院,吉林长春130062

出　　处：《中国生物化学与分子生物学报》2007年第7期554-559,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.30270973;30571335)~~

摘　　要：构建分别含有CMV、肌肉特异启动子SP及双启动子CMV-SP的真核报告基因EGFP表达载体(pCMV-EGFP、pSP-EGFP及pCMV-SP-EGFP)和生长激素释放因子(GRF)表达载体(pCMV-GRF、pSP-GRF和pCMV-SP-GRF).将3种EGFP表达质粒分别注射至小鼠骨骼肌.注射后1周、2周、3周以及4周时,分别提取肌肉组织RNA,应用半定量RT-PCR检测报告基因(EGFP)的表达量,发现pCMV-SP-EGFP组EGFP表达量显著高于pCMV-EGFP组和pSP-EGFP组(P<0.01);经荧光检测得到了较强的荧光信号.将3种GRF表达质粒分别注射至小鼠骨骼肌,在注射后每隔10d记录小鼠累积增重,采血并用RIA法测定血清胰岛素样生长因子Ⅰ(IGF-Ⅰ)浓度,结果pCMV-SP-GRF组累积增重、IGF-I浓度分别高于生理盐水组、pCMV-GRF组和pSP-GRF组(P<0.05).结果表明:CMV与SP两种启动子组合,在小鼠骨骼肌内使外源基因表达效率提高.本研究为提高外源基因在肌肉组织的表达提供了新的途径.Six vectors were constructed, which were carrying EGFP or GRF （pCMV-EGFP, pSP-EGFP, pCMV-SP-EGFP, pCMV-GRF, pSP-GRF and pCMV-SP-GRF） driven by CMV promoter, SP promoter and CMV-SP promoters, respectively. They were injected into skeletal muscle of mice to observe their expression efficacy. The differences of EGFP expressions of the vectors were evaluated by semi-quantitative polymerase chain reaction （PCR） at 1, 2, 3 and 4 weeks postinjection. The results showed that the expression of the EGFP with two promoters was significantly higher than the groups of pCMV-EGFP and pSP-EGFP （P 〈 0.01 ）. The accumulative weight gain of mice and concentrations of serum IGF-Ⅰwere measured in every 10 days. The results showed that injection of pCMV-SP-GRF lead to accumulative weight gain and serum IGF-Ⅰlevels were significantly higher than the groups of pCMV-GRF and pSP-GRF （ P 〈 0.05）. The results indicated that the two promoters combined gained more robust expression of exogenous gene. This result would provide a beneficial exploring to improve the efficiency of gene expression in skeletal muscle.

关 键 词：CMV启动子 SP启动子 双启动子 EGFP GRF IGF-Ⅰ 

分 类 号：Q786[生物学—分子生物学] S852.23[农业科学—基础兽医学]