文献类型：期刊文献

中文题名：逆转录病毒载体构建的hNT-3基因在大鼠成纤维细胞中的表达

英文题名：Stable Expression of Retroviral Constructed Gene of hNT-3 in Rat Primary Fibroblast Cells

作者：欧阳立明[1];周涛[2];薛冲[2];刘志敏[2];张嗣良[1]

机构：[1]华东理工大学生物工程学院,上海200237;[2]军事医学科学院生物工程研究所,北京100071

年份：2002

卷号：18

期号：1

起止页码：55

中文期刊名：中国生物化学与分子生物学报

外文期刊名：Chinese Journal of Biochemistry and Molecular Biology

收录：CSTPCD;;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；PubMed;

语种：中文

中文关键词：逆转录病毒载体;人神经营养因子-3;基因治疗;基因表达;大鼠成纤维细胞

外文关键词：retroviral vector, human neurotrophin 3(hNT 3), gene therapy

摘要：为了检验重组逆转录病毒对体外培养细胞的感染能力和效率 ,用逆转录病毒载体构建的人神经营养因子 3(pLXSN NT3)感染大鼠原代成纤维细胞 ,经G4 18筛选获得了稳定整合有外源hNT 3的工程细胞株 .RT PCR证实了外源基因hNT 3已整合到宿主细胞基因组 ,并可合成其mRNA ;PCR检测方法证明细胞株不含具有感染能力的病毒 ;Western印迹证明了细胞能正确表达hNT 3;大乳鼠背根神经节检测了细胞上清液中的NT 3生物活性 .
To test the transfection ability and effect of recombinant retroviral toward cells cultured in vitro , rat fibroblast cells were transfected by recombinant retrovirus pLXSN NT3 produced by recombinant package cells. Engineered cells were gained after G418 selection. RT PCR demonstrated the integration and mRNA synthesis of exogenous gene hNT 3, PCR verified there was no infectious retrovirus in the engineered cell clone. Western blot and rat root dorsal ganglia bioassay demonstrated the correctness and activity of NT 3 expressed by the engineered cells. The success of transfection in vitro made stable ground for further animal experiments of gene therapy.