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CLONING AND EXPRESSION OF HUMAN GTP CYCLOHYDROLASEⅠ(GCHⅠ)
Author(s): SU Yujin, JIANG Xiaohua, JIANG Zhongjian, ZHANG Yuxin, YANG Qiuhui, XU Qunyuan, YANG Hui
Pages: 349-
352
Year: 2003
Issue:
4
Journal: ACTA ANATOMICA SINICA
Keyword: 三磷酸鸟苷酸环化水解酶Ⅰ; RT-PCR; 原位杂交; 人胚肝脏;
Abstract: 目的研究人三磷酸鸟苷酸环化水解酶Ⅰ(GCHⅠ)基因在多巴胺代谢过程中的作用. 方法从人胚胎肝脏中提取总RNA,以RT-PCR法扩增GCHⅠ cDNA,克隆于PGEM-T-easy载体中,测序正确后再构建真核表达载体,转染猴肾成纤维细胞系COS7,原位杂交检测其表达. 结果 RT-PCR扩增出904bp的cDNA,并成功构建真核表达载体pCI-neo-GCHⅠ,原位杂交证实其在COS7表达阳性率为70%~80%. 结论 GCHⅠ有望用于帕金森病的基因治疗.
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