Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method
文献类型：期刊论文
| 作者 | An, Y; Ji, J; Wu, W; Lv, A; Huang, R; Xiu, Z; An, Y, Chinese Acad Sci, Inst Appl Ecol, Wenhua Rd 72, Shenyang 110016, Peoples R China |
| 刊名 | MOLECULAR BIOLOGY
 |
| 出版日期 | 2006 |
| 卷号 | 40期号:3页码:486-492 |
| 关键词 | Dna Shuffling Adomet Synthetase Dna Recombination Molecular Evolution Step |
| ISSN号 | 0026-8933 |
| 英文摘要 | We describe a new approach to in vitro DNA recombination termed the Separate-Mixing method in this study. The reaction process of this method consists of two stages: at the first stage the reaction was implemented in two parallel teams, which generated random recombination by template-switching of growing polynucleotides from primers in the presence of unidirectional single-stranded DNA fragments used as templates, and then both teams were mixed together for further extension and recombination of DNA sequences at the second stage. Due to this particular strategy, the reaction process was also accompanied by two other processes of DNA shuffling and StEP simultaneously. Two AdoMet synthetase genes, sam2 from Saccharomyces cerevisiae and metK from Escherichia coli, which have only 56% homology on the DNA level, were used for recombination with the Separate-Mixing method. DNA recombination was available after a single round of reaction. When 10 randomly selected recombinants were sequenced, an unshuffled parental clone was not found, nor was unexpected insertion, deletion, or rearrangement detected. An evolved gene, sam', was obtained after screening and selection, which could obviously increase the accumulation of AdoMet in S. cerevisiae. |
| 学科主题 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000238266900016 |
| 公开日期 | 2011-09-23 |
| 源URL | [http://210.72.129.5/handle/321005/54943]  |
| 专题 | 沈阳应用生态研究所_沈阳应用生态研究所 |
| 通讯作者 | An, Y, Chinese Acad Sci, Inst Appl Ecol, Wenhua Rd 72, Shenyang 110016, Peoples R China |
| 推荐引用方式 GB/T 7714 | An, Y,Ji, J,Wu, W,et al. Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method[J]. MOLECULAR BIOLOGY,2006,40(3):486-492. |
| APA | An, Y.,Ji, J.,Wu, W.,Lv, A.,Huang, R.,...&An, Y, Chinese Acad Sci, Inst Appl Ecol, Wenhua Rd 72, Shenyang 110016, Peoples R China.(2006).Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method.MOLECULAR BIOLOGY,40(3),486-492. |
| MLA | An, Y,et al."Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method".MOLECULAR BIOLOGY 40.3(2006):486-492. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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