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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
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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.
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
关键词 金属硫蛋白 诱变 化学诱变 CDNA 克隆 Metallothionein Sulfites Mutagenicity tests Ovary Cricetus
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