A general solution for opening double-stranded DNA for isothermal amplification

doi:10.1038/srep34582

	A general solution for opening double-stranded DNA for isothermal amplification
	Tang, Z (reprint author), Chinese Acad Sci, Chengdu Inst Biol, Nat Prod Res Ctr, Chengdu 610041, Peoples R China.; Tang, Z; Cui, X; Huang, X; Li, N; Yuan, Y; Dong, J; Chen, GY
刊名	SCIENTIFIC REPORTS
	2016
卷号	6 页码:34582（1-8）
DOI	10.1038/srep34582
产权排序	1
文献子类	Article
英文摘要	Nucleic acid amplification is the core technology of molecular biology and genetic engineering. Various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). However, most of these methods can only detect single stranded nucleic acid. Herein, we put forward a simple solution for opening double-stranded DNA for isothermal detection methods. The strategy employs recombination protein from E. coli (RecA) to form nucleoprotein complex with single-stranded DNA, which could scan double-stranded template for homologous sites. Then, the nucleoprotein can invade the double-stranded template to form heteroduplex in the presence of ATP, resulting in the strand exchange. The ATP regeneration system could be eliminated by using high concentration of ATP, and the 3'-OH terminal of the invasion strand can be recognized by other DNA modifying enzymes such as DNA polymerase or DNA ligase. Moreover, dATP was found to be a better cofactor for RecA, which make the system more compatible to DNA polymerase. The method described here is a general solution to open dsDNA, serving as a platform to develop more isothermal nucleic acids detection methods for real DNA samples based on it.
学科主题	Science & Technology - Other Topics
语种	英语
资助机构	National Sciences Foundation of China [21572222, 21402189, 21322208] ; National Sciences Foundation of China [21572222, 21402189, 21322208] ; National Sciences Foundation of China [21572222, 21402189, 21322208] ; National Sciences Foundation of China [21572222, 21402189, 21322208]
内容类型	期刊论文
源URL	[http://210.75.237.14/handle/351003/28178]
专题	成都生物研究所_天然产物研究
通讯作者	Tang, Z (reprint author), Chinese Acad Sci, Chengdu Inst Biol, Nat Prod Res Ctr, Chengdu 610041, Peoples R China.; Cui, X
推荐引用方式 GB/T 7714	Tang, Z ,Tang, Z,Cui, X,et al. A general solution for opening double-stranded DNA for isothermal amplification[J]. SCIENTIFIC REPORTS,2016,6:34582（1-8）.
APA	Tang, Z .,Tang, Z.,Cui, X.,Huang, X.,Li, N.,...&Chen, GY.(2016).A general solution for opening double-stranded DNA for isothermal amplification.SCIENTIFIC REPORTS,6,34582（1-8）.
MLA	Tang, Z ,et al."A general solution for opening double-stranded DNA for isothermal amplification".SCIENTIFIC REPORTS 6(2016):34582（1-8）.