心血管疾病基因治疗的基础研究Ⅰ．重组质粒pN2－LacZ在体外的表达  

BASIC RESEARCH OF GENE THE THERAPY ON CARDIOVASCULAR DISEASEI.EXPRESSION OF REPORT GENE LacZ IN VITRO

作　　者：姚阿卿 杨亚东[1] 朱小君[1] 李岱宗[1] 孙双丹 温进坤[1] 周爱儒[1] 汤健[1] 

机构地区：[1]北京医科大学心血管基础研究所分子生物学室

出　　处：《北京大学学报（医学版）》1994年第S1期111-114,共4页Journal of Peking University:Health Sciences

摘　　要：构建了带有标记基因ＬａｃＺ的逆转录病毒质粒ｐＮ２－ＬａｃＺ。用磷酸钙共沉淀的方法，将ｐＮ２－ＬａｃＺ转染培养的大鼠血管平滑肌细胞，Ｇ４１８筛选得到抗性集落。Ｓｏｕｔｈｅｒｎ和Ｎｏｒｔｈｅｒｎｂｌｏｔ分析表明，ＬａｃＺ基因已整合到细胞染色体上并转录出相应的ｍＲＮＡ。组化染色和酶活力测定结果表明，外源基因ＬａｃＺ可在血管平滑肌细胞中表达出有活性的蛋白质产物。提示，血管平滑肌细胞可作为心血管疾病基因治疗的靶细胞。o study whether a foreign gene can expression in cardiovascular cells,we constructed a retroviral plas-mid pN2-LacZ containing a report gene LacZ and a neomycin gene. Then this plasmid was transfected into rat vascular smooth muscle cells(VSMC)by coprecipitation with calcium phosphate. Many colonies of the VSMC were obtained by selected with G418.By means of Southern and Northern blot analysis,we found that the foreign gene LacZ was integrated on the chromosal DNA of VSMC and transcripted into mRNA in the transfected cells.The results of histochemical stain and enzyme activity assay demonstrated that the transfected VSMC can produce the active protein of LacZ gene.

关 键 词：标记基因LacZ 血管平滑肌细胞 基因治疗 

分 类 号：R459.9[医药卫生—治疗学]