基础医学与临床2011，Vol.31Issue(7)：740-745,6.
小鼠Islet-1基因慢病毒表达载体的构建及其诱导C3H10T1/2细胞向心肌样细胞特异性分化
Construction of lentiviral expression vector expressing mouse Islet-1 gene and investigation of Islet-1 mediated induction of Cs H1oT1/2 cell differentiation into cardiomyocyte-like cells
摘要
Abstract
Objective To construct lentiviral expression vectors targeting mouse gene Islet-1 and to study the effect of Islet-1 on stem cell differentiation. Methods Islet-1 gene was obtained by PCR and inserted into pLenO-WPI Vector. The positive plasmid was selected and infected 293T cells with helper plasmid to produce recombinaet lentivirus. Islet-1 expression and its related gene, gene and protein markers of heart, liver, bone, and nerves systems were measured by Real-time quantitative PCR and Western blot after C3H10T1/2 was infected by the lentiviruses. Results PCR and sequencing showed that the right DNA fragment and Islet-1 had been inserted and detected in both in gene and protein levels. Cardiac development related genes GATA-4, MEF2C; NKx2. 5 increased in 1 week and reached the highest level in 2 weeks, and cTnT reached a high level in 3 weeks and increasedover time. There were no expressions of gene and protein markers of liver, bone, brain. Conclusion Lentiviral expression vector carrying mouse lslet-1 gene was constructed. lslet-1 promoted C3 H10T1/2 cells differentiate into cardiomyocyte-like cells specifically, that laid the foundation for further investigation of Islet-1.关键词
慢病毒表达载体/Islet-1/干细胞特异性分化/心肌样细胞Key words
lentiviral expression vector/ lslet-1/ MSCS differentiation/ cardiomyocyte-like cells分类
生物科学引用本文复制引用
智深深,田杰,刘官信,鲁荣,林建萍,刘建平,朱静..小鼠Islet-1基因慢病毒表达载体的构建及其诱导C3H10T1/2细胞向心肌样细胞特异性分化[J].基础医学与临床,2011,31(7):740-745,6.基金项目
国家自然科学基金(30973219) （30973219）
自然科学基金重点项目(CSTC,2009BA5084) （CSTC,2009BA5084）
