大肠杆菌莽草酸途径限速酶多基因盒的构建及基因替换  

Construction of Rate-Limiting Multigene-Cassette in Shikimate Pathway and Gene Replacement in E.coli

作　　者：李永辉[1] 王世春[1] 刘云[1] 郭长江[2] 蒋与刚[2] 徐琪寿[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]军事医学科学院卫生学与环境医学研究所,天津300050

出　　处：《中国生物化学与分子生物学报》2004年第4期473-478,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：军事医学科学院创新基金 (No .990 2 5 0 2 ) ;天津市科技攻关项目 (No .0 3 3 182 711)~~

摘　　要：优化大肠杆菌芳香族氨基酸生物合成代谢途径 ,构建莽草酸代谢途径限速酶的多基因盒PtacaroAaroCaroBkan .利用Red重组系统 ,在破坏整体调控基因csrA时 ,替换多基因盒 .Southern印迹证实 ,基因破坏和基因替换是成功的 .摇瓶发酵表明 ,构建的基因工程菌株比原始菌株基础产酸率提高了 4 5Multigene cassette carrying Ptac, aroA, aroB, aroC and kan was constructed to optimize aromatic amino acids biosynthesis pathway in E.coli. The global regulatory gene csrA was disrupted and replaced by multigene cassette using Red recombinant system. Southern blotting confirmed that the gene disruption and replacement were successful. The flasks fermentation verified that the rate of conversion from glucose to amino acid of the modified strain increased 4.53 folds than that of the control.

关 键 词：多基因盒子 RED重组系统 基因破坏和基因替换 

分 类 号：Q786[生物学—分子生物学]