Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process

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	Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process
	An, YF; Ji, JF; Wu, WF; Huang, RB; Wei, YT; Xiu, ZL; Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China
刊名	BIOTECHNOLOGY LETTERS
	2008
卷号	30 期号:7 页码:1227-1232
关键词	Adomet Synthetase Directed Evolution Dna Recombination Error-prone Pcr S-adenosylmethionine
ISSN号	0141-5492
英文摘要	An efficient method for creating a DNA library is presented in which gene mutagenesis and recombination can be introduced by integrating error-prone PCR with a staggered extension process in one test tube. In this process, less than 15 cycles of error-prone PCR are used to introduce random mutations. After precipitated and washed with ethanol solution, the error-prone PCR product is directly used both as template and primers in the following staggered extension process to introduce DNA recombination. The method was validated by using adenosyl-methionine (AdoMet) synthetase gene, sam1, as a model. The full-length target DNA fragment was available after a single round. After being selected with a competitive inhibitor, ethionine, a mutated gene was obtained that increased AdoMet accumulation in vivo by 56%.
学科主题	Biotechnology & Applied Microbiology
语种	英语
WOS记录号	WOS:000256475900014
公开日期	2011-09-23
内容类型	期刊论文
源URL	[http://210.72.129.5/handle/321005/54425]
专题	沈阳应用生态研究所_沈阳应用生态研究所
通讯作者	Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China
推荐引用方式 GB/T 7714	An, YF,Ji, JF,Wu, WF,et al. Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process[J]. BIOTECHNOLOGY LETTERS,2008,30(7):1227-1232.
APA	An, YF.,Ji, JF.,Wu, WF.,Huang, RB.,Wei, YT.,...&Wu, WF, Chinese Acad Sci, Inst Appl Ecol, Shenyang, Peoples R China.(2008).Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process.BIOTECHNOLOGY LETTERS,30(7),1227-1232.
MLA	An, YF,et al."Random mutagenesis and recombination of sam1 gene by integrating error-prone PCR with staggered extension process".BIOTECHNOLOGY LETTERS 30.7(2008):1227-1232.