Cloning and identification of methionine synthase gene from Pichia pastoris
文献类型：期刊论文
| 作者 | Huang, L; Li, DY; Wang, SX; Zhang, SM; Chen, JH; Wu, XF |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2005 |
| 卷号 | 37期号:6页码:371-378 |
| 关键词 | methionine synthase Pichia pastoris rapid amplification of cDNA end (RACE) functional complementation |
| 通讯作者 | Wang, SX (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,shxwang@sibs.ac.cn |
| 英文摘要 | Methionine synthase (MS) is grouped into two classes. Class One MS (MetH) and Class Two MS (MetE) share no homology and differ in their catalytic model. Based on the conserved sequences of metE genes from different organisms, a segment of the metE gene was first cloned from Pichia pastoris genomic DNA by PCR, and its 5' and 3' regions were further cloned by 5'- and 3'-rapid amplification of cDNA ends (RACE), respectively. The assembled sequence reveals an open reading frame encoding a polypeptide of 768 residues, and the deduced product shares 76% identity with MetE of Saccharomyces cerevisiae. P. pastoris methionine synthase (PpMetE) consists of two domains common to MetEs. The active site is located in the C-terminal domain, in which the residues involved in the interaction of zinc with substrates are conserved. Homologous expression of PpMetE in P. pastoris was achieved, and the heterologous expression of PpMetE in the S. cerevisiae strain XJB3-1D that is Meff-defective restored the growth of the mutant on methionine-free minimal media. The gene sequence has been submitted to GenBank/EMBL/DDBJ under accession No. AY601648. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | SACCHAROMYCES-CEREVISIAE ; ESCHERICHIA-COLI ; HOMOCYSTEINE ; ZINC |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000229992500002 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1925]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Huang, L,Li, DY,Wang, SX,et al. Cloning and identification of methionine synthase gene from Pichia pastoris[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2005,37(6):371-378. |
| APA | Huang, L,Li, DY,Wang, SX,Zhang, SM,Chen, JH,&Wu, XF.(2005).Cloning and identification of methionine synthase gene from Pichia pastoris.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,37(6),371-378. |
| MLA | Huang, L,et al."Cloning and identification of methionine synthase gene from Pichia pastoris".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 37.6(2005):371-378. |
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