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| 人PSCA真核表达载体的构建及表达 | |
| 引用本文： | 任杰,高江平,阎瑾琦,肖毅,王伟,王宇,江乐,于继云.人PSCA真核表达载体的构建及表达[J].军医进修学院学报,2010,31(6):607-609. |
| 作者姓名： | 任杰 高江平 阎瑾琦 肖毅 王伟 王宇 江乐 于继云 |
| 作者单位： | 1. 军事医学科学院,基础医学研究所,北京,100850;解放军总医院,泌尿外科,北京,100853 2. 解放军总医院,泌尿外科,北京,100853 3. 军事医学科学院,基础医学研究所,北京,100850 |
| 基金项目： | 国家“863”基金项目(2006AA02A237,2007AA02Z451);;国家自然科学基金项目(30772002,30840094)~~ |
| 摘 要： | 目的构建含有信号肽及FLAG标签的人前列腺干细胞抗原（PSCA）真核表达载体,并在人胚肾293T细胞中进行表达。方法通过PCR方法获得SIG-FLAG基因片段及人PSCA基因片段,插入到真核表达载体pIRES-neo中。构建的重组质粒pIRES-neo-sig-FLAG-PSCA转染293T细胞,利用流式细胞仪、免疫荧光及RT-PCR方法检测其表达情况。结果PCR扩增出的SIG-FLAG及PSCA基因测序正确,酶切鉴定证明重组质粒pIRES-neo-sig-FLAG-PSCA构建成功;检测结果显示重组质粒pIRES-neo-sig-FLAG-PSCA在293T细胞中得到表达。结论成功构建了重组质粒pIRES-neo-sig-FLAG-PSCA,且在293T细胞中能有效表达,为后续转人PSCA基因细胞系的构建工作奠定了基础。 |
| 关 键 词： | 前列腺 干细胞 抗原 基因表达 293T细胞 |
Construction of eukaryotic expression vector for human prostate stem cell antigen and its expression |
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| REN Jie,GAO Jiang-ping,YAN Jin-qi,XIAO Yi,WANG Wei,WANG Yu,JIANG Le,YU Ji-yun.Construction of eukaryotic expression vector for human prostate stem cell antigen and its expression[J].Academic Journal of Pla Postgraduate Medical School,2010,31(6):607-609. | |
| Authors: | REN Jie GAO Jiang-ping YAN Jin-qi XIAO Yi WANG Wei WANG Yu JIANG Le YU Ji-yun |
| Institution: | REN Jie1,2,GAO Jiang-ping2,YAN Jin-qi1,XIAO Yi1,WANG Wei1,WANG Yu1,JIANG Le1,YU Ji-yun1 1 Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing 100850,China,2Department of Urology,Chinese PLA General Hospital,Beijing 100853 |
| Abstract: | Objective To construct the eukaryotic expression plasmid of human prostate stem cell antigen(PSCA)containing a signal peptide and a FLAG tag and to detect its expression in eukaryotic 293T cells.Methods SIG-FLAG and human PSCA gene was constructed by PCR and inserted into a eukaryotic expression vector pIRES-neo.The recombinant plasmid pIRES-neo-sig-FLAG-PSCA was tansfected to the 293T cells,and its expression was detected by FACS,IMF and RT-PCR,respectively.Results The gene sequence of SIG-FLAG and human P... |
| Keywords: | Prostate Stem Cell Antigen Gene Expression 293T Cells |
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