| 米根霉L-乳酸脱氢酶基因的克隆及在大肠杆菌中的表达 |
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| 引用本文： | 郑志,姜绍通,章建国,罗水忠,潘丽军,蒋诗平. 米根霉L-乳酸脱氢酶基因的克隆及在大肠杆菌中的表达[J]. 合肥工业大学学报(自然科学版), 2006, 29(4): 489-491 |
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| 作者姓名： | 郑志 姜绍通 章建国 罗水忠 潘丽军 蒋诗平 |
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| 作者单位： | 合肥工业大学,生物与食品工程学院,安徽,合肥,230009;中国科学技术大学,同步辐射国家重点实验室,安徽,合肥,230029 |
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| 基金项目： | 高等学校博士学科点专项科研项目 , 合肥工业大学校科研和教改项目 , 教育部研究生创新基金 |
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| 摘 要： | 文章以米根霉基因组DNA为模板,根据已公布的米根酶L-乳酸脱氢酶基因(ldnL)序列设计引物,PCR扩增得到含有ldnL的DNA片段;PCR产物T/A克隆后再双酶切,将ldnL片段连接到大肠杆菌表达载体pET17b,得到重组表达质粒pET17b-ldnL;pET17b-ldnL转化大肠杆菌BL21(DE3),摇瓶培养诱导表达后的菌体经SDS-PAGE电泳分析,结果表明克隆的ldnL基因在大肠杆菌中实现表达。
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| 关 键 词： | L-乳酸脱氢酶基因 大肠杆菌 克隆 |
| 文章编号： | 1003-5060(2006)04-0489-03 |
| 修稿时间： | 2004-12-16 |
Cloning of L-lactate dehydrogenase gene(ldnL) from Rhizopus oryzae and its expression in Escherichia coli |
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| ZHENG Zhi,JIANG Shao-tong,ZHANG Jian-guo,LUO Shui-zhong,PAN Li-jun,JIANG Shi-ping. Cloning of L-lactate dehydrogenase gene(ldnL) from Rhizopus oryzae and its expression in Escherichia coli[J]. Journal of Hefei University of Technology(Natural Science), 2006, 29(4): 489-491 |
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| Authors: | ZHENG Zhi JIANG Shao-tong ZHANG Jian-guo LUO Shui-zhong PAN Li-jun JIANG Shi-ping |
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| Abstract: | A fragment of L-lactate dehydrogenase gene(ldnL) was cloned from the genomic DNA of Rhizopus oryzae by polymerase chain reaction(PCR) based on the released ldnL sequence.The PCR product was double-digested after T/A clone and the digested fragment was ligated with pET17b,a kind of expression vector in Escherichia coli(E.coli).Then,the recombinant plasmid,pET17b-ldnL,was transformed into the host cell E.coli BL21(DE3).The transformed cell was measured by SDS-PAGE after culturing in the inducing medium and the result showed that the cloned ldnL was expressed in BL21(DE3). |
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| Keywords: | L-lactate dehydrogenase gene(ldnL) Escherichia coli cloning |
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