家畜生态学报2013，Vol.34Issue(2)：15-18,4.
人血小板生成素基因的克隆及CHO稳定细胞系的建立
Cloning of Human Thrombopoietin Gene and Establishment of CHO Cell Lines Stably Expressing TPO Gene
摘要
Abstract
Human thrombopoietin (Human Thrombopoietin, hTPO) gene was cloned by PCR using the human liver cDNA as template, then fragment of hTPO gene was cloned into pcDNA3. 1 ( + ) eukaryotic expression vector which has the neomycin resistance gene (neo) selection marker, named as pcDNA3. 1 ( + )-hTPO. First, the recombinant plasmid was transiently transfected into 293T cells. The transient expression of TPO in eukaryotic cells was determined by Western blot using mouse anti-human TPO monoclonal antibody; then Chinese hamster ovary cells (Chinese hamster ovary, CHO) were transfected with pcDNA3. 1 ( + ) -hTPO recombinant plasmid, using G418 of 400 jug/ml to select CHO stable cell lines, and verified by PCR and Western blot. The above results showed that three CHO cell lines stably expressing TPO gene were established. The cell lines may get a large number of proteins applied to actual medical and activity functional experiments and even laid a certain foundation for the next stage.关键词
人血小板生成素/基因克隆/CHO稳定细胞系/Western blotKey words
human thrombopoietin/ gene clone/ CHO stable cell lines/ Western blot分类
农业科技引用本文复制引用
马倩倩,宋玲玲,王红梅,方永志,王立群,何洪彬..人血小板生成素基因的克隆及CHO稳定细胞系的建立[J].家畜生态学报,2013,34(2):15-18,4.基金项目
泰山学者海外特聘专家专项 （）
国家奶牛产业技术体系建设专项 （）
国家自然科学基金(31272586) （31272586）
国家转基因重大专项(2009ZX08007-006B,2011ZX08007-002,2011ZX08008-004) （2009ZX08007-006B,2011ZX08007-002,2011ZX08008-004）
济南市高校院所主创新计划(201004027,201202059,201102034) （201004027,201202059,201102034）
