Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures
文献类型：期刊论文
| 作者 | Wang, Jing1,3; Zhu, Jianming3; Wang, Chenru3; Zhou, Guangqing3; Yu, Xin3; Fan, Huijun3; An, Ran2,3; Komiyama, Makoto3; Liang, Xingguo2,3 |
| 刊名 | BIOCHEMISTRY
 |
| 出版日期 | 2020-02-04 |
| 卷号 | 59期号:4页码:400-406 |
| ISSN号 | 0006-2960 |
| DOI | 10.1021/acs.biochem.9b00866 |
| 通讯作者 | Komiyama, Makoto(xgsz@ouc.edu.cn) ; Liang, Xingguo(liangxg@ouc.edu.cn) |
| 英文摘要 | Thermus thermophilus DNA ligase (Tth DNA ligase) is widely employed for cloning, enzymatic synthesis, and molecular diagnostics at high temperatures (e.g., 65 degrees C). It has been long believed that the complementary ends must be very long (e.g., >30 bp) to place two DNA fragments nearby for the ligation. In the current study, the length of the complementary portion was systematically varied, and the ligation efficiency was evaluated using the high resolution melting (HRM) method. Unexpectedly, very short oligonucleotides (7-10 nt) were successfully ligated on the complementary overhang attached to a dsDNA at 70 degrees C. Furthermore, sticky ends with the overhang of only 4 nt long, available after scission with many restriction enzymes, were also efficiently ligated at 45-70 degrees C. The ligation yield for the 6-nt-long sticky ends was as high as 80%. It was concluded that Tth DNA ligase can be used as a unique tool for DNA manipulation that cannot be otherwise easily accomplished. |
| 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 |
| 语种 | 英语 |
| WOS记录号 | WOS:000511509600008 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/165352]  |
| 专题 | 中国科学院海洋研究所 |
| 通讯作者 | Komiyama, Makoto; Liang, Xingguo |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Qingdao, Peoples R China 3.Ocean Univ China, Qingdao, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Jing,Zhu, Jianming,Wang, Chenru,et al. Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures[J]. BIOCHEMISTRY,2020,59(4):400-406. |
| APA | Wang, Jing.,Zhu, Jianming.,Wang, Chenru.,Zhou, Guangqing.,Yu, Xin.,...&Liang, Xingguo.(2020).Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures.BIOCHEMISTRY,59(4),400-406. |
| MLA | Wang, Jing,et al."Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures".BIOCHEMISTRY 59.4(2020):400-406. |
入库方式： OAI收割
来源：海洋研究所
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