dc.contributor.author | 王生育 | zh_CN |
dc.contributor.author | 颜江华 | zh_CN |
dc.contributor.author | 潘阳霖 | zh_CN |
dc.contributor.author | 李学军 | zh_CN |
dc.contributor.author | 陈忠 | zh_CN |
dc.date.accessioned | 2011-04-26T08:24:06Z | |
dc.date.available | 2011-04-26T08:24:06Z | |
dc.date.issued | 2008-04-25 | zh_CN |
dc.identifier.citation | 生物工程学报 Chinese Journal of Biotechnology,200824(4):553-557 | zh_CN |
dc.identifier.issn | 1000-3061 | zh_CN |
dc.identifier.uri | https://dspace.xmu.edu.cn/handle/2288/8488 | |
dc.description.abstract | 从GenBank获得人PTP1B催化活性区(PTP1Bc)氨基酸序列(1~301aa),通过重叠PCR获得PTP1Bc基因。构建pET-22b(+)/PTP1Bc原核表达载体,转化大肠杆菌BL21(DE3),阳性重组子IPTG诱导表达,Ni柱纯化蛋白。目的蛋白以包涵体的形式表达,表达量占菌体总蛋白30%以上。纯化后,蛋白纯度达95%以上。Western blotting结果表明所得的蛋白可与抗PTP1B抗体发生特异性结合;酶活实验证实复性的蛋白具有一定的磷酸酶活性。PTP1Bc基因的构建、表达纯化及活性分析,为进一步的功能研究奠定了基础。
【英文摘要】 The amino acid sequence (1-301aa) coding the human PTP1B catalytic domain (PTP1Bc) was obtained from the GenBank.The PTP1Bc gene was constructed by overlapping PCR,then was inserted into vector pET-22b(+) and expressed efficiently in E.coli BL21(DE3) under optimum condition after IPTG induction.The proteins were expressed mainly as inclusion bodies with the yield of more than 30% of total bacterial proteins.The expressed products were purified through Ni~(2+)-affinity chromatographic column.After purificati... | zh_CN |
dc.description.sponsorship | 厦门市重大疾病攻关研究基金(No.3502220051027);; 卫生部(福建省)卫生教育联合攻关项目(No.Wkj2005-2-019)资助~~ | zh_CN |
dc.language.iso | zh | zh_CN |
dc.publisher | 中国科学院微生物所;中国微生物学会 | zh_CN |
dc.subject | PTP1Bc | zh_CN |
dc.subject | 基因表达 | zh_CN |
dc.subject | 蛋白纯化 | zh_CN |
dc.subject | 活性分析 | zh_CN |
dc.subject | expression | zh_CN |
dc.subject | protein purification | zh_CN |
dc.subject | activity analysis | zh_CN |
dc.title | 蛋白酪氨酸磷酸酶PTP1B催化活性区的原核表达及活性分析 | zh_CN |
dc.title.alternative | Expression and Activity Analysis of Catalytic Domain of PTP1B | zh_CN |
dc.type | Article | zh_CN |