Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis
文献类型：期刊论文
| 作者 | Zhu, Jianming3; Wang, Jing1,3; Cheng, Kai3; Chen, Hui3; An, Ran3; Zhang, Yaping3; Komiyama, Makoto3; Liang, Xingguo2,3 |
| 刊名 | CHEMBIOCHEM
 |
| 出版日期 | 2019-12-05 |
| 页码 | 5 |
| ISSN号 | 1439-4227 |
| 关键词 | enzyme catalysis high-resolution melting kinetics ligases quantitative analysis |
| DOI | 10.1002/cbic.201900489 |
| 通讯作者 | Wang, Jing(wangjing2019@qdio.ac.cn) ; Liang, Xingguo(liangxg@ouc.edu.cn) |
| 英文摘要 | High-resolution melting (HRM) analysis has been improved and applied for the first time to quantitative analysis of enzymatic reactions. By using the relative ratios of peak intensities of substrates and products, the quantitativity of conventional HRM analysis has been improved to allow detailed kinetic analysis. As an example, the ligation of sticky ends through the action of T4 DNA ligase has been kinetically analyzed, with comprehensive data on substrate specificity and other properties having been obtained. For the first time, the kinetic parameters (k(obs) and apparent K-m) of sticky-end ligation were obtained for both fully matched and mismatched sticky ends. The effect of ATP concentration on sticky-end ligation was also investigated. The improved HRM method should also be applicable to versatile DNA-transforming enzymes, because the only requirement is that the products have T-m values different enough from the substrates. |
| 资助项目 | National Natural Science Foundation of China[31571937] ; Shandong Provincial Natural Science Foundation of China[ZR2019BC096] ; Fundamental Research Funds for the Central Universities[201812009] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| WOS记录号 | WOS:000500756500001 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/163822]  |
| 专题 | 海洋研究所_海洋生态与环境科学重点实验室 |
| 通讯作者 | Wang, Jing; Liang, Xingguo |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Marine Ecol & Environm Sci, Qingdao 266071, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Drugs & Bioprod, Qingdao 266003, Peoples R China 3.Ocean Univ China, Coll Food Sci & Engn, Qingdao 266003, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhu, Jianming,Wang, Jing,Cheng, Kai,et al. Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis[J]. CHEMBIOCHEM,2019:5. |
| APA | Zhu, Jianming.,Wang, Jing.,Cheng, Kai.,Chen, Hui.,An, Ran.,...&Liang, Xingguo.(2019).Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis.CHEMBIOCHEM,5. |
| MLA | Zhu, Jianming,et al."Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis".CHEMBIOCHEM (2019):5. |
入库方式： OAI收割
来源：海洋研究所
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