Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method

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	Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method
	An, Y ; Ji, J ; Wu, W ; Lv, A ; Huang, R ; Xiu, Z
刊名	MOLECULAR BIOLOGY
	2006
卷号	40 期号:3 页码:486-492
关键词	DNA shuffling AdoMet synthetase DNA recombination molecular evolution StEP
ISSN号	0026-8933
通讯作者	An, Y, Chinese Acad Sci, Inst Appl Ecol, Wenhua Rd 72, Shenyang 110016, Peoples R China
中文摘要	We describe a new approach to in vitro DNA recombination termed the Separate-Mixing method in this study. The reaction process of this method consists of two stages: at the first stage the reaction was implemented in two parallel teams, which generated random recombination by template-switching of growing polynucleotides from primers in the presence of unidirectional single-stranded DNA fragments used as templates, and then both teams were mixed together for further extension and recombination of DNA sequences at the second stage. Due to this particular strategy, the reaction process was also accompanied by two other processes of DNA shuffling and StEP simultaneously. Two AdoMet synthetase genes, sam2 from Saccharomyces cerevisiae and metK from Escherichia coli, which have only 56% homology on the DNA level, were used for recombination with the Separate-Mixing method. DNA recombination was available after a single round of reaction. When 10 randomly selected recombinants were sequenced, an unshuffled parental clone was not found, nor was unexpected insertion, deletion, or rearrangement detected. An evolved gene, sam', was obtained after screening and selection, which could obviously increase the accumulation of AdoMet in S. cerevisiae.
学科主题	Biochemistry & Molecular Biology
收录类别	SCI
语种	英语
WOS记录号	WOS:000238266900016
公开日期	2011-09-23
内容类型	期刊论文
源URL	[http://210.72.129.5/handle/321005/54943]
专题	沈阳应用生态研究所_沈阳应用生态研究所_期刊论文
推荐引用方式 GB/T 7714	An, Y,Ji, J,Wu, W,et al. Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method[J]. MOLECULAR BIOLOGY,2006,40(3):486-492.
APA	An, Y,Ji, J,Wu, W,Lv, A,Huang, R,&Xiu, Z.(2006).Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method.MOLECULAR BIOLOGY,40(3),486-492.
MLA	An, Y,et al."Molecular evolution of adomet synthetase by DNA recombination with a novel separate-mixing method".MOLECULAR BIOLOGY 40.3(2006):486-492.