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心血管疾病基因治疗的基础研究Ⅲ.重组质粒fPGV-MT-LacZ在大鼠血管平滑肌细胞中的表达

BASIC RESEARCH OF GENE THERAPY ON CARDIOVASCULAR DISEASE Ⅲ.EXPRESSION OF REPORT GENE LacZ IN RAT VAXCULAR SMOOTH MUSCLE MUSCLE CELLS

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【作者】 朱小君胡征毕万里唐朝枢周爱儒汤健

【Author】 Institute of Cardiovascular Research Zhu Xiaojun,et al

【机构】 北京医科大学心血管基础研究所分子生物学室

【摘要】 为观察外源性基因在心血管细胞内的表达,本工作构建了含有外源性标记基因LacZ的逆转录病毒质粒fPGV-MT-LacZ。应用脂质体(Lipofectin)包裹质粒,转染培养的大鼠主动脉平滑肌细胞,经G4l8筛选、克隆以及β-半乳糖苷酶的酶活测定,证明外源性基因可以在血管平滑肌细胞中表达。揭示血管平滑肌细胞可作为心血管疾病基因治疗的一种靶细胞。

【Abstract】 retroviral plasmid fPGV-MT-LacZ containing a report gene LacZ and a neomycin gene is constructed and transfected to rat vascular smooth muscle cells(VSMC )by means of lipofectin. Genetic engineered VSMC are obtained by selected with G4l8.By means of Southern and Northern blot analvsis,we find that the foreign gene LacZ is integrated on the chromosal DNA of the VSMC and transcripted into mRNA. The cells can produce β-galactosidase and become blue due to adding X-gal.Using β-galactosidase assay system,the activity of β-galactosidase in the transfected VSMC is 2.3 × 10-4U/107 cells. These datas suggested that the foreign gene LacZ can express in the rat vascular smooth muscle cells.

  • 【文献出处】 北京医科大学学报 ,JOURNAL OF BEIJING MEDICAL UNIVERSITY , 编辑部邮箱 ,1994年S1期
  • 【分类号】R459.9
  • 【下载频次】64
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