Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures

doi:10.1021/acs.biochem.9b00866

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	Thermus thermophilus DNA Ligase Connects Two Fragments Having Exceptionally Short Complementary Termini at High Temperatures
	Wang, Jing 1,3; Zhu, Jianming 3; Wang, Chenru 3; Zhou, Guangqing 3; Yu, Xin 3; Fan, Huijun 3; An, Ran 2,3; Komiyama, Makoto 3; Liang, Xingguo 2,3
刊名	BIOCHEMISTRY
	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
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000511509600008
内容类型	期刊论文
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.