| 大肠杆菌巴豆甜菜碱还原酶基因缺失菌株的构建 |
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| 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),2004,30(5):519-522. |
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| 作者姓名： | 张文刚 叶江 吴海珍 张惠展 |
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| 作者单位： | 华东理工大学生物反应器工程国家重点实验室,上海,200237 |
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| 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。
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| 关 键 词： | 同源重组 巴豆甜菜碱还原酶 基因缺失 抗性基因 突变株 |
| 文章编号： | 1006-3080(2004)05-0519-04 |
| 修稿时间： | 2003年11月5日 |
Deletion of Crotonobetaine Reductase Gene in Escherica coli |
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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,2004,30(5):519-522. |
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| Authors: | ZHANG Wen-gang YE Jiang WU Hai-zhen ZHANG Hui-zhan |
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| Institution: | ZHANG Wen-gang,YE Jiang,WU Hai-zhen,ZHANG Hui-zhan~* |
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| 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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| Keywords: | homologous recombination crotonobetaine reductase gene deletion resistant gene mutantstrain |
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