A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR
文献类型：期刊论文
| 作者 | An, YF; Ji, JF; Wu, WF; Lv, A; Huang, RB; Wei, YT; Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2005 |
| 卷号 | 68期号:6页码:774-778 |
| ISSN号 | 0175-7598 |
| 英文摘要 | A rapid and efficient method to perform site-directed mutagenesis based on an improved version of overlap extension by polymerase chain reaction (OE-PCR) is demonstrated in this paper. For this method, which we name modified (M)OE-PCR, there are five steps: (1) synthesis of individual DNA fragments of interest (with average 20-bp overlap between adjacent fragments) by PCR with high-fidelity pfu DNA polymerase, (2) double-mixing (every two adjacent fragments are mixed to implement OE-PCR without primers), (3) pre-extension (the teams above are mixed to obtain full-length reassembled DNA by OE-PCR without primers), (4) synthesis of the entire DNA of interest by PCR with outermost primers and template DNA from step 3, (5) post-extension (ten cycles of PCR at 72 degrees C for annealing and extension are implemented). The method is rapid, simple and error-free. It provides an efficient choice, especially for multiple-site mutagenesis of DNAs; and it can theoretically be applied to the modification of any DNA fragment. Using the MOE-PCR method, we have successfully obtained a modified sam1 gene with eight rare codons optimized simultaneously. |
| 学科主题 | Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000232856700009 |
| 公开日期 | 2011-09-23 |
| 源URL | [http://210.72.129.5/handle/321005/55053]  |
| 专题 | 沈阳应用生态研究所_沈阳应用生态研究所 |
| 通讯作者 | Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China |
| 推荐引用方式 GB/T 7714 | An, YF,Ji, JF,Wu, WF,et al. A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2005,68(6):774-778. |
| APA | An, YF.,Ji, JF.,Wu, WF.,Lv, A.,Huang, RB.,...&Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China.(2005).A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,68(6),774-778. |
| MLA | An, YF,et al."A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 68.6(2005):774-778. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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