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目的将含有connexin 43基因的质粒在脂质体的介导下转染进入大鼠L6骨骼肌成肌细胞,使用G418根据有限稀释法筛选单克隆细胞株L6myoblast-Cx43。方法通过免疫细胞化学技术、Western blot技术,对整合有connexin43基因的阳性细胞克隆进行检测。结果两个阳性克隆组Cx43蛋白表达均明显高于对照组。结论获得的两个持续高表达connexin43蛋白的细胞株可用于进行移植细胞之间及移植细胞与宿主细胞之间能否形成良好的电偶联的研究。
OBJECTIVE: The plasmid containing connexin 43 gene was transfected into rat L6 skeletal muscle myoblasts by liposome, and the monoclonal cell line L6myoblast-Cx43 was selected by limiting dilution method with G418. Methods Immunocytochemistry and Western blot were used to detect the positive cell clones with connexin43 gene. Results The expression of Cx43 protein in two positive clones was significantly higher than that in the control group. Conclusions The two cell lines with continuously high expression of connexin43 protein can be used to study whether there is a good electrical coupling between the transplanted cells and between the transplanted cells and the host cells.