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    张文刚, 叶江, 吴海珍, 张惠展. 大肠杆菌巴豆甜菜碱还原酶基因缺失菌株的构建[J]. 华东理工大学学报（自然科学版）, 2004, (5): 519-522.
    引用本文: 张文刚, 叶江, 吴海珍, 张惠展. 大肠杆菌巴豆甜菜碱还原酶基因缺失菌株的构建[J]. 华东理工大学学报（自然科学版）, 2004, (5): 519-522.
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    ZHANG Wen-gang, YE Jiang, WU Hai-zhen, ZHANG Hui-zhan~*. Deletion of Crotonobetaine Reductase Gene in Escherica coli[J]. Journal of East China University of Science and Technology, 2004, (5): 519-522.
    Citation: ZHANG Wen-gang, YE Jiang, WU Hai-zhen, ZHANG Hui-zhan~*. Deletion of Crotonobetaine Reductase Gene in Escherica coli[J]. Journal of East China University of Science and Technology, 2004, (5): 519-522.
    shu

    大肠杆菌巴豆甜菜碱还原酶基因缺失菌株的构建

    Deletion of Crotonobetaine Reductase Gene in Escherica coli

      摘要
    • 摘要: 报道了体外构建caiA基因缺失的带有卡那霉素抗性基因的5．2kb线状DNA分子，以此转化大肠杆菌JM83和BL21(DE3)株，借助于体内DNA同源重组，定向敲除了大肠杆菌中的巴豆甜菜碱还原酶编码基因caiA。经遗传稳定性实验、聚合酶链反应(PCR)以及Southern鉴定，表明所获得的JM83转化子22号和BL21(DE3)转化子4号确为caiA基因缺失突变株；酶活分析结果表明，22号和4号转化子均丧失了巴豆甜菜碱还原酶活性。

       

      Abstract: This work reports that the construction of a 5.2 kb DNA fragment in which the caiA gene was replaced by kanamycin resistant gene(Km~r), which was then transferred into JM83 and BL21(DE3). The No.4 and 22 strains from BL21(DE3) and JM83 respectively, were proved to be caiA-defective through the experiment of genetic stability, polymerase chain reaction(PCR) and southern blotting; and enzyme assays indicated that strains 4 and 22 have lost their crotonobetaine reductase activities.

       

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