中国仓鼠卵巢MTⅡ　cDNA的体外诱变及其变种克隆的筛选  

MUTAGENESIS OF CHINESE HAMSTER OVARY CELL METALLOTHIONEIN-Ⅱ cDNA IN VITRO AND SCREENING OF ITS MUTANT CLONES

作　　者：张建业[1,2] 庞维秋[1,2] 张莲英[1,2] 王鑫[1,2] 胡晓燕[1,2] 吴伟芳[1,2] 

机构地区：[1]山东医科大学生物化学教研室 [2]山东医科大学卫生统计学教研室

出　　处：《山东医科大学学报》1996年第4期275-278,共4页Acta Academiae Medicinae Shandong

基　　金：山东省自然科学基金

摘　　要：用亚硫酸钠对中国仓鼠卵细胞金属硫蛋白Ⅱ（ＣＨＯＭＴⅡ）的编码序列进行体外诱变，将变异ＤＮＡ在大肠杆菌ＢＤ１５２８（ｕｎｇ－）中扩增，使ＭＴⅡ编码序列中ＣＧ碱基对被ＴＡ碱基对置换。从１０６个ＭＴⅡ变种克隆中发现了１２个新变种，其中１０个变种的变异导致了１～３个氨基酸残基的改变。１７个点突变中的１１个使半胱氨酸转变成酪氨酸。这些变异可能对ＭＴⅡ结合金属的能力和抗原性等生物学性质产生影响。The Chinese hamster ovary cell metallothionein-Ⅱ(CHO MT-Ⅱ ) coding sequence was mutagenized by sodium bisulfite in vitro and the treated DNA was propagated in E. coli BD1528(ung ),which result in replacement of CG base pair with TA base pair finally. 12 new mutants were found from 106 MT-Ⅱ mutant clones. Mutation in each of 10 new mutants will cause one to three amino acid residues to be changed. Eleven of seventeen point mutations will lead to replacement of cysteine with tyrosine. Those mutations will possibly produce certain effect on biological properties of MT-Ⅱ, such as metal-binding capacity and antigenicity of the protein and so on.

关 键 词：金属硫蛋白 诱变 化学诱变 CDNA 克隆 

分 类 号：Q513[生物学—生物化学] R394－33[医药卫生—医学遗传学]