A convenient and robust method for construction of combinatorial and random mutant libraries
文献类型：期刊论文
| 作者 | An, Yingfeng; Wu, Wenfang; Lv, Anguo |
| 刊名 | BIOCHIMIE
 |
| 出版日期 | 2010-08-01 |
| 卷号 | 92期号:8页码:1081-1084 |
| 关键词 | Mutant Library Dna Shuffling Error-prone Pcr Ligase-independent Cloning |
| ISSN号 | 0300-9084 |
| DOI | 10.1016/j.biochi.2010.03.016 |
| 英文摘要 | Here we describe a convenient and robust ligase-independent method for construction of combinatorial and random mutant libraries. The homologous genes flanked by plasmid-derived DNA sequences are fragmented, and the random fragments are reassembled in a self-priming polymerase reaction to obtain chimeric genes. The product is then mixed with linearized vector and two pairs of flanking primers, followed by assembly of the chimeric genes and linearized vector by PCR to introduce recombinant plasmids of a combinatorial library. Commonly, it is difficult to find proper restriction sites during the construction of recombinant plasmids after DNA shuffling with multiple homologous genes. However, this disadvantage can be overcome by using the ligase-independent method because the steps of DNA digestion and ligation can be avoided during library construction. Similarly, DNA sequences with random mutations introduced by error-prone PCR can be used to construct recombinant plasmids of a random mutant library with this method. Additionally, this method can meet the needs of large and comprehensive DNA library construction. (C) 2010 Elsevier Masson SAS. All rights reserved. |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000280570800021 |
| 出版者 | ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER |
| 源URL | [http://210.72.129.5/handle/321005/121591]  |
| 专题 | 中国科学院沈阳应用生态研究所 |
| 通讯作者 | Wu, Wenfang |
| 作者单位 | Chinese Acad Sci, Inst Appl Ecol, Shenyang 110016, Peoples R China |
| 推荐引用方式 GB/T 7714 | An, Yingfeng,Wu, Wenfang,Lv, Anguo. A convenient and robust method for construction of combinatorial and random mutant libraries[J]. BIOCHIMIE,2010,92(8):1081-1084. |
| APA | An, Yingfeng,Wu, Wenfang,&Lv, Anguo.(2010).A convenient and robust method for construction of combinatorial and random mutant libraries.BIOCHIMIE,92(8),1081-1084. |
| MLA | An, Yingfeng,et al."A convenient and robust method for construction of combinatorial and random mutant libraries".BIOCHIMIE 92.8(2010):1081-1084. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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