| 鼠李糖乳杆菌D-(+)-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： | 黄艳燕,黄靖华,卢福芝,孙靓,彭立新,周兴,黄日波.鼠李糖乳杆菌D-(+)-乳酸脱氢酶基因的克隆及在大肠杆菌中的表达[J].生物技术通报,2010(12). |
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| 作者姓名： | 黄艳燕 黄靖华 卢福芝 孙靓 彭立新 周兴 黄日波 |
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| 作者单位： | 1. 广西科学院国家非粮生物质能源工程技术研究中心,南宁,530003
2. 广西大学,南宁,530003 |
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| 基金项目： | 国家科技支撑计划项目(2007BAD75B06); 广西科学基金项目(桂科攻0782003-4) |
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| 摘 要： | 利用PCR的方法从鼠李糖乳杆菌基因组DNA中扩增到D-(+)-乳酸脱氢酶基因(ldhD),并连接到载体pSE380上,构建表达质粒pSE-ldhD,将重组质粒pSE-ldhD转化大肠杆菌BL21(DE3),重组菌株经IPTG诱导表达,SDS-PAGE电泳分析表明ldhD在大肠杆菌中实现了表达,表达产物的分子量约为37kD。同时采用紫外分光光度法测定D-乳酸脱氢酶的酶活,测得重组菌株的D-乳酸脱氢酶活力为5.4U/mL,最适反应温度为35℃,最适pH为5.6。
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| 关 键 词： | D-( )-乳酸脱氢酶 克隆 表达 鼠李糖乳杆菌 |
Cloning of D-(+)-lactate Dehydrogenase Gene from Lactobacillus rhamnosus and Expression in Escherichia coli |
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| Huang Yanyan,Huang Jinghua,Lu Fuzhi,Sun Liang,Peng Lixin,Zhou Xing,Huang Ribo.Cloning of D-(+)-lactate Dehydrogenase Gene from Lactobacillus rhamnosus and Expression in Escherichia coli[J].Biotechnology Bulletin,2010(12). |
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| Authors: | Huang Yanyan Huang Jinghua Lu Fuzhi Sun Liang Peng Lixin Zhou Xing Huang Ribo |
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| Institution: | Huang Yanyan1 Huang Jinghua2 Lu Fuzhi2 Sun Liang1 Peng Lixin1 Zhou Xing1 Huang Ribo1(1Guangxi Academy of Sciences,National Engineering Research Center for Non-foodBiorefinery,Nanning 530003,2Guangxi University,Nanning 530003) |
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| Abstract: | The gene ldhD encoding D-(+)-lactate dehydrogenase was amplified from genome DNA of Lactobacillus rhamnosus by PCR.The PCR product was ligated with the expression vector pSE380 after double-digested.A recombinant plasmid pSE-ldhD was constructed and transformed into E.coli BL21(DE3).The D-(+)-lactate dehydrogenase activity of recombinant strain was determined with spectrum assay after IPTG induction.SDS-PAGE result showed that the cloned ldhD was expressed in the E.coli,and the relative molecular weight of ... |
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| Keywords: | D-( )-lactate dehydrogenase Clone Expression Lactobacillus rhamnosus |
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