| 人癌症/睾丸相关蛋白XAGE-1b原核表达载体的构建及融合蛋白的表达 |
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| 引用本文： | 郑慧玲,;刘振杰,;陈碧,;丁航,;刘新光.人癌症/睾丸相关蛋白XAGE-1b原核表达载体的构建及融合蛋白的表达[J].湛江医学院学报,2014(4):432-434. |
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| 作者姓名： | 郑慧玲 ;刘振杰 ;陈碧 ;丁航 ;刘新光 |
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| 作者单位： | [1]广东省医学分子诊断重点实验室,广东东莞523808; [2]广东省中医院检验科,广东广州510370; [3]广东医学院附属医院,广东湛江524001; [4]广东医学院生物化学与分子生物学研究所,广东湛江524023 |
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| 基金项目： | 国家自然科学基金项目（No.81170327）; 广东省自然科学基金（No.05011579）; 东莞市科技计划项目（No.2012108102022）; 广东医学院科研基金（No.XQ0621） |
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| 摘 要： | 目的构建人癌症/睾丸相关蛋白XAGE-1b原核表达载体及诱导XAGE-1融合蛋白表达。方法从文库质粒pACT2-XAGE-1b中利用PCR法扩增XAGE-b cDNA编码序列,将其克隆到pET-32a载体中,并将重组质粒pET-32bXAGE-1b转染大肠杆菌DE3中,用异丙基硫代半乳糖苷（IPTG）诱导表达,Western blot检测融合蛋白表达。结果原核表达载体pET-32a-XAGE-1b构建成功;转染菌株经IPTG诱导后,出现约29 kD蛋白过表达,Western-blot检测显示His6融合蛋白表达。结论成功构建原核表达载体pET-32a-XAGE-1b,转染菌株可表达His6融合蛋白。
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| 关 键 词： | XAGE-1b 原核表达 融合蛋白 |
Construction and fusion protein expression of prokaryotic expression vector of human cancer/testis related protein XAGE-1b |
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| Institution: | ZHENG Hui-ling, LIU Zhen-jie, CHEN Bi, DING Hang, LIU Xin-guang (1. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China) |
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| Abstract: | Objective To construct the prokaryotic expression vector of human cancer/testis related protein XAGE-1b and induce XAGE-1 fusion protein expression. Methods The XAGE-1b cDNA was amplified by PCR from pACT2-XAGE-1b and inserted into expression vector pET-32 b.The recombinant plasmid pET-32b-XAGE-1b was transfected into ？DE3 strain of Escheria coli, followed by the induction of isopropyl a-D-1-thiogalactopyranoside（IPTG）. XAGE-1 fusion protein expression was detected by Western blot. Results The prokaryotic expression vector pET-32b-XAGE-1b was successfully constructed. After induced by IPTG, the transfection strain showed overexpression of approximately 29 kD protein, and Western-blot revealed His6 fusion protein expression. Conclusion The prokaryotic expression vector pET-32b-XAGE-1b transfected strain can express His6 fusion protein. |
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| Keywords: | XAGE-1b prokaryotic expression fusion protein |
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