Institutional Repository of Key Laboratory of Marine Ecology & Environmental Sciences, CAS
| Effective Characterization of DNA Ligation Kinetics by High-Resolution Melting Analysis | |
| Zhu, Jianming1; Wang, Jing1,3; Cheng, Kai1; Chen, Hui1; An, Ran1; Zhang, Yaping1; Komiyama, Makoto1; Liang, Xingguo1,2 | |
| 2019-12-05 | |
| Source Publication | CHEMBIOCHEM
 |
| ISSN | 1439-4227 |
| Pages | 5 |
| Abstract | 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. |
| Keyword | enzyme catalysis high-resolution melting kinetics ligases quantitative analysis |
| DOI | 10.1002/cbic.201900489 |
| Indexed By | SCI |
| Language | 英语 |
| Funding Project | Fundamental Research Funds for the Central Universities[201812009] ; Shandong Provincial Natural Science Foundation of China[ZR2019BC096] ; National Natural Science Foundation of China[31571937] ; National Natural Science Foundation of China[31571937] ; Shandong Provincial Natural Science Foundation of China[ZR2019BC096] ; Fundamental Research Funds for the Central Universities[201812009] |
| WOS Research Area | Biochemistry & Molecular Biology ; Pharmacology & Pharmacy |
| WOS Subject | Biochemistry & Molecular Biology ; Chemistry, Medicinal |
| WOS ID | WOS:000500756500001 |
| Publisher | WILEY-V C H VERLAG GMBH |
| Citation statistics | |
| Document Type | 期刊论文 |
| Version | 出版稿 |
| Identifier | http://ir.qdio.ac.cn/handle/337002/163823 |
| Collection | 海洋生态与环境科学重点实验室 |
| Corresponding Author | Wang, Jing; Liang, Xingguo |
| Affiliation | 1.Ocean Univ China, Coll Food Sci & Engn, Qingdao 266003, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Drugs & Bioprod, Qingdao 266003, Peoples R China 3.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Marine Ecol & Environm Sci, Qingdao 266071, Peoples R China |
| Corresponding Author Affilication | Key Laboratory of Marine Ecology & Environmental Sciences, CAS |
| Recommended Citation 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. |
| Files in This Item: | ||||||
| File Name/Size | DocType | Version | Access | License | ||
| Effective Characteri（833KB） | 期刊论文 | 作者接受稿 | 开放获取 | CC BY-NC-SA | View | |
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