%0 Journal Article
%T Studies on the Two Methods of Gene Random Mutation in vitro<br>基因体外随机突变的两种方法的研究
%A LI Hong-jian
%A ZHANG Yi
%A ZHOU Tian-hong
%A LI Yue-qin
%A <br>李弘剑
%A 张毅
%A 周天鸿
%A 李月琴
%J 遗传
%D 2000
%I 
%X Two novel and simple methods were described in the paper for in vitro mutagenesis and recombinatin of polynucleotide sequence to mimic and accelerate nature's recombination strategy to direct the evolution of protein function.One is chemical mutagenesis: protein gene was inserted into M13mp18,and the single-stranded DNA was treated with 1 0mol/L sodium nitrite at room temperature for 1h for mutation and converted into duplex,then the mutated gene was ligated to plasmid for expression.Another is elongation mutagenesis: random peptide of 10 amino acid was connected at C terminal of protein to expand the sequence space by increasing protein's dimensions,then the elongation mutated gene expressed in E.coli.We have used these two methods to recombine the thermostabilized catalase I, and these two methods were found to be efficient to form a lot of catalase I mutates by identifying the properties of mutated enzyme.
%K directed evolution
%K chemical mutagenesis
%K elongation mutagenesis
%K catalase I
%K Bacillusstearothermophilus<br>定向进化
%K 化学突变
%K 延伸突变
%K 蛋白质
%K 随机突变
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=3E23F50E071DF18E21B2F5AEE4F1FA5E&aid=EC3EA726FC9BBED0&yid=9806D0D4EAA9BED3&vid=BC12EA701C895178&iid=0B39A22176CE99FB&sid=6700D0D256586E73&eid=8C83C265AD318E34&journal_id=0253-9772&journal_name=遗传&referenced_num=4&reference_num=4