| 弗氏柠檬酸菌甘油脱水酶基因在大肠杆菌中的克隆和表达 |
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| 引用本文： | 齐向辉,朱绮霞,韦宇拓,黄鲲,黄日波.弗氏柠檬酸菌甘油脱水酶基因在大肠杆菌中的克隆和表达[J].工业微生物,2005,35(3):10-13. |
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| 作者姓名： | 齐向辉 朱绮霞 韦宇拓 黄鲲 黄日波 |
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| 作者单位： | 1. 广西大学广西亚热带生物资源保护利用重点实验室,南宁,530005
2. 南宁中诺生物工程有限公司,南宁,530005 |
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| 基金项目： | 国家“863”资助项目（编号：2003AA001039） |
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| 摘 要： | 以弗氏柠檬酸菌（Citrobacter freundii）基因组DNA为模板，通过PCR得到甘油脱水酶（glycerol dehydratase）基因dhaB、dhaC、dhaE，克隆到表达载体pSE380上，得到重组质粒pSn-dhaBCE。将此重组质粒转化到E．coli JM109中，重组菌株SDS-PAGE结果显示有明显的61kD、22kD、16kD三条特异性蛋白条带出现。重组菌株经诱导表达，酶活力为11．59U／mL。
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| 关 键 词： | 弗氏柠檬酸菌 甘油脱水酶 克隆 表达 克隆和表达 酶基因 柠檬酸 大肠杆菌 SDS-PAGE 基因组DNA |
Gene cloning and expression of glycerol dehydratase from Citrobacter freundii in Escherichia coli |
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| QI Xiang-hui,ZHU Qi-xia,WEI Yu-tuo,HUANG Kun,HUANG Ri-bo.Gene cloning and expression of glycerol dehydratase from Citrobacter freundii in Escherichia coli[J].Industrial Microbiology,2005,35(3):10-13. |
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| Authors: | QI Xiang-hui ZHU Qi-xia WEI Yu-tuo HUANG Kun HUANG Ri-bo |
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| Abstract: | The genes dhaB, dhaC, dhaE encoding glycerol dehydratase of Citrobacter freundii were amplified by PCR. The recombinant plasmid pSE-dhaBCE was constructed by inserting dhaB, dhaC, dhaE genes imo expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaBCE. Glycerol dehydratase was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The deduced products of the dhaBCE genes with calculated molecular masses of 61kDa, 22kDa, 16kDa, respectively, revealed high activity. By comparison, no activity appeared in hosted E. coli JM109. |
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| Keywords: | Citrobacter freundii glycerol dehydratase cloning expression |
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