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 1; Wang, Chenru 1; Zhou, Guangqing 1; Yu, Xin 1; Fan, Huijun 1; An, Ran 1,2; Komiyama, Makoto 1; Liang, Xingguo 1,2
	2020-02-04
Source Publication	BIOCHEMISTRY
ISSN	0006-2960
Volume	59 Issue:4 Pages:400-406
Corresponding Author	Komiyama, Makoto(xgsz@ouc.edu.cn) ; Liang, Xingguo(liangxg@ouc.edu.cn)
Abstract	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.
DOI	10.1021/acs.biochem.9b00866
Indexed By	SCI
Language	英语
Funding Project	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
WOS Subject	Biochemistry & Molecular Biology
WOS ID	WOS:000511509600008
Publisher	AMER CHEMICAL SOC
Citation statistics
Document Type	期刊论文
Identifier	http://ir.qdio.ac.cn/handle/337002/165353
Collection	中国科学院海洋研究所
Corresponding Author	Komiyama, Makoto; Liang, Xingguo
Affiliation	1.Ocean Univ China, Qingdao, Peoples R China 2.Qingdao Natl Lab Marine Sci & Technol, Qingdao, Peoples R China 3.Chinese Acad Sci, Inst Oceanol, Qingdao, Peoples R China
First Author Affilication	Institute of Oceanology, Chinese Academy of Sciences
Recommended Citation 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.

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