Construction of a high-efficiency multi-site-directed mutagenesis
文献类型：期刊论文
| 作者 | Tan, Haidong2; Li, Yueguang3; Chen, Ling1; Kasai, Tomonari1; Seno, Masaharu1 |
| 刊名 | african journal of biotechnology
 |
| 出版日期 | 2011-01-17 |
| 卷号 | 10期号:3页码:449-452 |
| 关键词 | Site-directed multibase mutagenesis shift primer hexapeptide gene enterokinase gene |
| ISSN号 | 待补充 |
| 产权排序 | 1,1 |
| 通讯作者 | masaharuseno |
| 中文摘要 | construction of a high-efficiency multi-site-directed mutagenesis |
| 英文摘要 | although site-directed mutagenesis has been used in many fields, it still has low rate of success and high cost because of low-yield target products. a modified method for multi-site-directed mutagenesis was developed with shifted primer design and cold-start polymerase chain reaction (pcr). the developed method was successfully applied to hexapeptide gene synthesis and recombinant enterokinase gene modification in the plasmids pet41a and pet24b-ek. the efficiency was pronounced at a 1:10 molar ratio of 7-base mutant products to 705-bp fragment products as control. even in a 10-base substitution mutagenic pcr, a 1:50 molar ratio of mutant products to 705-bp fragment products was reached. meanwhile, the quality of mutants was proved through the transformation efficiency and sequencing. this method was beneficial to prepare high-quality multibase mutagenesis and also implied that large-scale multibase mutagenesis was feasible, efficient, economical, and productive. |
| WOS标题词 | science & technology ; life sciences & biomedicine |
| 学科主题 | 物理化学 |
| 类目[WOS] | biotechnology & applied microbiology |
| 研究领域[WOS] | biotechnology & applied microbiology |
| 关键词[WOS] | pcr |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000286397500022 |
| 公开日期 | 2012-07-09 |
| 源URL | [http://159.226.238.44/handle/321008/115580]  |
| 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.Okayama Univ, Dept Med & Bioengn Sci, Grad Sch Nat Sci & Technol, Okayama 7008530, Japan 2.Chinese Acad Sci, Dalian Inst Chem Phys, Dept Biotechnol, Dalian 116023, Peoples R China 3.Tianjin 4th Ctr Hosp, Dept Gen Surg, Tianjin 300140, Peoples R China |
| 推荐引用方式 GB/T 7714 | Tan, Haidong,Li, Yueguang,Chen, Ling,et al. Construction of a high-efficiency multi-site-directed mutagenesis[J]. african journal of biotechnology,2011,10(3):449-452. |
| APA | Tan, Haidong,Li, Yueguang,Chen, Ling,Kasai, Tomonari,&Seno, Masaharu.(2011).Construction of a high-efficiency multi-site-directed mutagenesis.african journal of biotechnology,10(3),449-452. |
| MLA | Tan, Haidong,et al."Construction of a high-efficiency multi-site-directed mutagenesis".african journal of biotechnology 10.3(2011):449-452. |
入库方式： OAI收割
来源：大连化学物理研究所
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