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酪氨酸羟化酶基因载体-pCMVTH 质粒的构建及在大鼠骨骼肌内的表达
引用本文:杨慧,孙阳,田竟生,郑少鹏,张进禄,赵春礼,徐群渊.酪氨酸羟化酶基因载体-pCMVTH 质粒的构建及在大鼠骨骼肌内的表达[J].中国组织化学与细胞化学杂志,1995(4).
作者姓名:杨慧  孙阳  田竟生  郑少鹏  张进禄  赵春礼  徐群渊
作者单位:首都医科大学神经科学研究所
摘    要:为用转基因方法治疗巴金森氏病大鼠模型,本研究采用分子克隆技术,将合成多巴胺的关键酶-酪氨酸羟化酶(TH)的基因,克隆进入以巨细胞病毒CMV为启动子的载体质粒内,经限制性内切酶定位分析证实该重组的DNA质粒的可靠性。携带TH基因的PCMVTH质粒以LIPO-FECTIN介导,在培养的原代骨骼肌细胞中高效表达。本研究为进一步用转基因的细胞植入脑内以治疗巴金森氏病打下一定基础。

关 键 词:酪氨酸羟化酶,DNA质粒重组,原代骨骼肌细胞,大鼠巴金森氏病

RECONSTRUCTION OF TYROSINE HYDROXYLASE GENE VECTOR-pCMVTH AND EXPRESSION IN RAT MUSCLE CELLS
Yang Hui, Sun Yang,Tia Jingsheng, Zheng Shaopeng,Zhang Jinlu, Zhao Chunli, Xu Qunyuan.RECONSTRUCTION OF TYROSINE HYDROXYLASE GENE VECTOR-pCMVTH AND EXPRESSION IN RAT MUSCLE CELLS[J].Chinese Journal of Histochemistry and Cytochemistry,1995(4).
Authors:Yang Hui  Sun Yang  Tia Jingsheng  Zheng Shaopeng  Zhang Jinlu  Zhao Chunli  Xu Qunyuan
Abstract:The purpose of this study was to determine whether the plasmid pCMVTH which reconstructed was able to show expression in rat muscle cells. We used tyrosine hydroxylasegene, which was from the plasmid of pKSTH as a transgene and cytomegalovirus (CMV),which was from the plasmid of pCMVLacZ as a cellular promoter to do subcloning. Theplasmid pCMVTH was identified by analysis of restriction endonuclease. Primary rat muscleCells were trans feeted with pCMVTH by lipofectin. About 30% myoblasts and myotubeshaving been cultured for 2 days showed expression of tyrosine hydroxylase gene wiht immunohistochmical staining. Our results may serve as a basis for in gene therapy inParkinson's disease by tranferring tyrosine hydroxylase gene into brain tissues.
Keywords:Tyrosine hydroxylase gene  pCMVTH  Primary muscle cells  RatParkinson's disease model  
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