PCR-based strategy for construction of multi-site-saturation mutagenic expression library
文献类型：期刊论文
| 作者 | Wang, Jinxia1,2; Zhang, Sufang1; Tan, Haidong1; Zhao, Zongbao (Kent)1 |
| 刊名 | journal of microbiological methods
 |
| 出版日期 | 2007-12-01 |
| 卷号 | 71期号:3页码:225-230 |
| 关键词 | expression library PCR cloning protein engineering site-saturation mutagenesis |
| 英文摘要 | there is an increasing demand for efficient and effective methods to engineer protein variants for industrial applications, structural biology and drug development. we describe a pcr-based strategy that produces multi-site-saturation mutagenic expression library using a circular plasmid carrying the wild-type gene. this restriction digestion- and ligation-independent method involves three steps: 1) synthesis of the degenerate oligonucleotide primers, 2) incorporation of the mutations through pcr, 3) transformation into the expression host. our strategy is demonstrated through successful construction of an e coli k12 malic enzyme expression library that contains members with simultaneous mutations on amino acid residues g311, d345 and g397. this method is in principle compatible with any circular vector that can be propagated with a dam(+) e. coli host to generate protein variant library with multiple changes, including mutation, short sequence deletion and insertion, or any mix of them. (c) 2007 elsevier b.v all rights reserved. |
| WOS标题词 | science & technology ; life sciences & biomedicine |
| 类目[WOS] | biochemical research methods ; microbiology |
| 研究领域[WOS] | biochemistry & molecular biology ; microbiology |
| 关键词[WOS] | nad-malic enzyme ; directed evolution ; cloning ; design |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000252080300007 |
| 公开日期 | 2015-11-17 |
| 源URL | [http://159.226.238.44/handle/321008/140594]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 作者单位 | 1.Dalian Inst Chem Phys, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Jinxia,Zhang, Sufang,Tan, Haidong,et al. PCR-based strategy for construction of multi-site-saturation mutagenic expression library[J]. journal of microbiological methods,2007,71(3):225-230. |
| APA | Wang, Jinxia,Zhang, Sufang,Tan, Haidong,&Zhao, Zongbao .(2007).PCR-based strategy for construction of multi-site-saturation mutagenic expression library.journal of microbiological methods,71(3),225-230. |
| MLA | Wang, Jinxia,et al."PCR-based strategy for construction of multi-site-saturation mutagenic expression library".journal of microbiological methods 71.3(2007):225-230. |
入库方式： OAI收割
来源：大连化学物理研究所
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