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马铃薯脂氢过氧化物裂解酶cDNA的克隆及其在大肠杆菌中的表达 被引量：1

Cloning of Hydroperoxide Lyase cDNA from Potato's Leaves and Its Expression in Escherichia coli
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摘要 采用反转录聚合酶链式反应(RT-PCR),以马铃薯幼叶总RNA为模板,扩增出1443bp的脂氢过氧化物裂解酶(HPL)cDNA(GenBank登录号GQ281583),将该基因片段克隆到表达载体PET-22b(+)上并转化到E.coliBL21(DE3),得到重组菌E.coliBL21(PET-22b-HPL),以0.5mmol/LIPTG诱导该重组菌过量表达目的蛋白(HPL),SDS-PAGE显示,重组HPL分子量为54ku。经紫外分光光度法检测,重组HPL的比酶活约为0.12U/mL。 The total RNA of potato＇ s immature leaves was isolated by one step method with Trizol reagent. Then 1443-bp cDNA （GenBank GQ281583） of hydroperoxide lyase was amplified by RT-PCR. The target gene was inserted into the expression vector PET-22b（ ＋ ） , which was finally expressed in Escherichia coli BL21 （DE3） by IPTG induction. SDS-PAGE analysis showed that the relative molicular mass of hydroperoxide lyase was about 54 ku. The specific activity of recomebinant hydroperoxide lyase was about 0. 12 U/mL.
作者 薛庆海 沐万孟 江波 华欲飞
机构地区 江南大学食品科学与技术国家重点实验室
出处 《食品与发酵工业》 CAS CSCD 北大核心 2009年第12期34-38,共5页 Food and Fermentation Industries
基金 科技部"863"计划资助项目(2008AA10Z305) 江南大学食品学院青年博士科研创新开放基金(项目号FS-200806)
关键词 马铃薯 脂氢过氧化物裂解酶 CDNA 克隆 表达 potato hydroperoxide lyase, cDNA, cloning, expression
分类号 S532 [农业科学—作物学]
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食品与发酵工业
2009年 第12期

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