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Expression of human cardiac troponin I-C in E-coli and its purification
Yan, CL; Xu, GB; Xia, TA
关键词Cardiac Troponin I-c Fusion Protein Expression Purification
刊名PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS
2004-09-01
31期:9页:812-817
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]Biochemistry & Molecular Biology ; Biophysics
英文摘要

In order to clone and express human cardiac troponin I-C fusion protein in application to the quality control for cTnI detection system, human cardiac troponin I and troponin C cDNAs were amplified from human cardiac using gene-specific primers designed from the published cDNA sequences by the polymerase chain reaction. The full-length of cTnI was linked with TnC by a short DNA sequence coding for 19 neutral amino acid residues. An expression construct for cTnI-C was engineered by inserting the corresponding cDNA into a pET15b plasmid. Then recombinant plasmid was transformed into E. coli BL21 (DE3)pLysS cells, and protein expression was induced by isopropyl-beta-D-thiogalactopyranoside (IPTG). Soluable expression of eTnI-C in prokaryotic system was successfully obtained. Fusion protein had an N-terminal His-tag sequence which could be purified by affinity chromatography on a Ni2+-Sepharose column. After one step affinity chromatography the fusion protein shows homogeneity as judged by SDS-PAGE. The fusion protein was stable and easy to be purified, could be used as candidate reference material for cTnI detection systems.

语种中文
WOS记录号WOS:000224052600008
引用统计
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/50209
专题北京大学第一临床医学院
作者单位Peking Univ, Hosp 1, Dept Lab Med Sci, Beijing 100034, Peoples R China
推荐引用方式
GB/T 7714
Yan, CL,Xu, GB,Xia, TA. Expression of human cardiac troponin I-C in E-coli and its purification[J]. PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,2004,31(9):812-817.
APA Yan, CL,Xu, GB,&Xia, TA.(2004).Expression of human cardiac troponin I-C in E-coli and its purification.PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS,31(9),812-817.
MLA Yan, CL,et al."Expression of human cardiac troponin I-C in E-coli and its purification".PROGRESS IN BIOCHEMISTRY AND BIOPHYSICS 31.9(2004):812-817.
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