摘要
目的将携带人降钙素(CT)基因的真核表达载体pcDNA3.0-hCT转染L-6成肌细胞,检测该基因在成肌细胞中的表达,为下一步基因治疗奠定基础。方法利用L ipofectam ineTM2000转染pcDNA3.0-hCT到L-6成肌细胞中;经G418筛选成功后,用逆转录—聚合酶链反应(RT-PCR)法检测人降钙素基因的表达,并检测细胞外分泌情况。结果从分子水平上成功检测到降钙素基因的表达;并且在培养液中检测到了平均42.22ng/106/24h浓度降钙素分泌。结论成功转染人降钙素基因真核表达载体pcDNA3.0-hCT于成肌细胞中,并可以在其中持续稳定地表达。
Objective To transform the expressing vector of human calcitonin （CT） gene, pcDNA3.0 - hCT, into the L6 myoblast and to identify the express of CT in L6 myoblast in order to make foundations for the next gene therapy. Methods The pcDNA3.0 -hCT was transformed into L6 myoblast. After successful filtration by G418, the expression of CT was identified and determined by RT - PCR and radioimmunoassay. Resuits At the level of molecule, we have identified the expression of hCT; in the cell culture medium, we have determined average density of hCT, which was 42.22ng/10^6/24h. Conclusion The pcDNA3.0 - hCT was successfully transformed into L6 myoblast and could express CT sustainedly.
出处
《中原医刊》
2006年第7期1-3,共3页
Central Plains Medical Journal
基金
国家自然科学基金资助项目(项目号:30271316)
