军事医学2013，Vol.37Issue(5)：354-357,4.DOI:10.7644/j.issn.1674-9960.2013.05.009
人G250真核表达载体pIRES-neo-G250的构建及表达
Construction and expression of recombinant plasmid pIRES-neo-G250 expressing human G250
李云奇 1张巍 2于继云 2高江平 2朱晓明 1肖毅 2王伟 2李盼 2王宇 2王琳 2徐元基 2阎瑾琦3
作者信息
- 1. 解放军总医院泌尿外科,北京,100853
- 2. 军事医学科学院基础医学研究所,北京,100850
- 3. 解放军316医院,北京,100093
- 折叠
摘要
Abstract
Objective To construct the eukaryotic expression plasmid pIRES -neo-G250 which expresses human G250, transduce it into the 293T cell and detect its expression. Methods The human G250 gene was amplified by PCR and linked into PMD18T,which was then inserted into the eukaryotic expression vector pIRES -neo. The constructed recombinant plasmid pIRES-neo-G250 was tansfected into the 293T cells , and its expression was determined by FACS , IMF and RT-PCR. Results The gene sequence of human G 250 which was amplified by PCR was right. Enzyme digestion analysis re -vealed that the recombinant plasmid pIRES -neo-G250 was successfully constructed. FACS, IMF and RT-PCR revealed that the recombinant plasmid was expressed in 293T cells. Conclusion The successful construction of recombinant plasmid pIRES-neo-G250 and its effective expression in 293T cells can serve further construction of human G250-transfected cell line.关键词
G250/真核表达/质粒/293T细胞/肾肿瘤Key words
G250 / eukaryotic expression/ plasmids / 293T cell/ kidney neoplasms分类
医药卫生引用本文复制引用
李云奇,张巍,于继云,高江平,朱晓明,肖毅,王伟,李盼,王宇,王琳,徐元基,阎瑾琦..人G250真核表达载体pIRES-neo-G250的构建及表达[J].军事医学,2013,37(5):354-357,4.
