变性双链法实现葡萄糖异构酶基因敲除的研究  

Glucose Isomerasc Gene Knock-out by Denatured Double-stranded DNA

作　　者：廖军 徐冲[1] 杨永辉 李晖 程阳 陈承露 朱国萍 牛立文[3] 王玉珍 

机构地区：[1]中国科学技术大学生命科学学院分子生物与细胞生物学系,合肥230027 [2]国科学技术大学生命科学学院分子生物与细胞生物学系,合肥230027 [3]国科学技术大学结构生物学开放实验室,合肥230027

出　　处：《Acta Genetica Sinica》2000年第5期449-454,共6页

基　　金：国家"八六三"高技术计划!( 130-13-02-04)

摘　　要：对７号淀粉酶菌Ｍ１０３３的遗传背景进行分析，建立了其原生质体制备、转化的优化条件。构建了葡萄糖异构酶（ＧＩ）结构基因内插入硫链丝菌肽基因（ｔｓｒ）的置换型同源重组质粒，利用其变性双链片段实现与Ｍ１０３３菌株染色体上基因的同源重组，获得置换型葡萄糖异构酶缺陷型菌株Ｍ１０３３ＬＪ。为在染色体上引入突变位点实现染色体上分子定点改造奠定了基础。After Genetic background analysis of Streptomyces diastaticus No. 7 strain M1033, the modified conditions of M1033 protoplasts and transformation were established. Replacement plasmid for homologous recombination was also constructed by inserting tsr gene into glucose isomerase gene. The homologous recombination of GI gene in M1033 chromosomes was achieved by using denatured linearized DNA fragments and glucose isomerase deficient strain M1033LJ was obtained. It is basic for introducing mutation into M1033 chromosome and realizing site-directed molecular reformation.

关 键 词：葡萄糖异构酶 同源重组 变性双链法 基因敲除 

分 类 号：Q754[生物学—分子生物学]