Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process
文献类型：期刊论文
| 作者 | An, YF ; Ji, JF ; Wu, WF ; Huang, RB ; Wei, YT ; Xiu, ZL |
| 刊名 | BIOTECHNOLOGY LETTERS
 |
| 出版日期 | 2008 |
| 卷号 | 30期号:7页码:1227-1232 |
| 关键词 | AdoMet synthetase directed evolution DNA recombination error-prone PCR S-adenosylmethionine |
| ISSN号 | 0141-5492 |
| 通讯作者 | Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China |
| 中文摘要 | An efficient method for creating a DNA library is presented in which gene mutagenesis and recombination can be introduced by integrating error-prone PCR with a staggered extension process in one test tube. In this process, less than 15 cycles of error-prone PCR are used to introduce random mutations. After precipitated and washed with ethanol solution, the error-prone PCR product is directly used both as template and primers in the following staggered extension process to introduce DNA recombination. The method was validated by using adenosyl-methionine (AdoMet) synthetase gene, sam1, as a model. The full-length target DNA fragment was available after a single round. After being selected with a competitive inhibitor, ethionine, a mutated gene was obtained that increased AdoMet accumulation in vivo by 56%. |
| 学科主题 | Biotechnology & Applied Microbiology |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000256475900014 |
| 公开日期 | 2011-09-23 |
| 源URL | [http://210.72.129.5/handle/321005/54425]  |
| 专题 | 沈阳应用生态研究所_沈阳应用生态研究所_期刊论文 |
| 推荐引用方式 GB/T 7714 | An, YF,Ji, JF,Wu, WF,et al. Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process[J]. BIOTECHNOLOGY LETTERS,2008,30(7):1227-1232. |
| APA | An, YF,Ji, JF,Wu, WF,Huang, RB,Wei, YT,&Xiu, ZL.(2008).Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process.BIOTECHNOLOGY LETTERS,30(7),1227-1232. |
| MLA | An, YF,et al."Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process".BIOTECHNOLOGY LETTERS 30.7(2008):1227-1232. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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