Expression, purification and activity of human myristoyl-CoA: N-myristoyltransferase
文献类型：期刊论文
| 作者 | Yang, XY; Chen, L; Chen, H; Xu, ZP; Li, BL |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 1999-03 |
| 卷号 | 31期号:2页码:175-179 |
| 关键词 | human NMT PCR gene expression His(6) fusion expression metal (Ni2+) chelation affinity chromatography |
| ISSN号 | 1672-9145 |
| 文献子类 | Article |
| 英文摘要 | A gene encoding human myristoyl-CoA: protein N-myristoyltransferase (hNMT) from a brain cDNA library has been obtained by PCR amplification and DNA sequencing. Then the mature-type and His(6)-fusion-type expression plasmids (pMF-hNMT(3) and pMFHT-hNMT(2)) containing the hNMT gene under control of T7 promoter have been constructed and transformed into E. coli BL21(DE3). SDS-PAGE analysis showed that the recombinant hNMT products expressed at 37 degrees C were almost unsoluble, but most of the His(6)-hNMT product expressed at lower temperature (22 degrees C) was soluble and its yield was about 7% of the total soluble cellular proteins. By immobilized metal (Ni2+) chelation affinity chromatography, up to 80% His(6)-hNMT was purified by one step from bacterial lysate. The labelling experiment in vitro demonstrated that the expressed and purified His(6)-hNMT had an obvious catalytic activity to transferring myristoyl group. |
| WOS关键词 | SACCHAROMYCES-CEREVISIAE ; GENE |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000079339100015 |
| 出版者 | OXFORD UNIV PRESS |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/274744]  |
| 专题 | 中国科学院上海药物研究所 |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Biochem, Shanghai 200031, Peoples R China 2.Zhejiang Univ, Dept Biol Sci & Technol, Hangzhou 310027, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 200031, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yang, XY,Chen, L,Chen, H,et al. Expression, purification and activity of human myristoyl-CoA: N-myristoyltransferase[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,1999,31(2):175-179. |
| APA | Yang, XY,Chen, L,Chen, H,Xu, ZP,&Li, BL.(1999).Expression, purification and activity of human myristoyl-CoA: N-myristoyltransferase.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,31(2),175-179. |
| MLA | Yang, XY,et al."Expression, purification and activity of human myristoyl-CoA: N-myristoyltransferase".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 31.2(1999):175-179. |
入库方式： OAI收割
来源：上海药物研究所
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