| 川西高原黑唇鼠兔乳酸脱氢酶C基因的原核表达 |
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| 引用本文： | 张庆莲,刘浩浩,贺庆华,刘东奇,李蒙,梁正旭.川西高原黑唇鼠兔乳酸脱氢酶C基因的原核表达[J].安徽农业科学,2011,39(14):8622-8623,8626. |
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| 作者姓名： | 张庆莲 刘浩浩 贺庆华 刘东奇 李蒙 梁正旭 |
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| 作者单位： | 成都医学院检验医学院临床生化教研室;西南民族大学生命科学与技术学院;四川省阿坝县畜牧兽医局; |
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| 基金项目： | 四川省应用基础研究(2008JY0068); 国家自然科学基金(31071700); 国家民委资助课题(08XN04) |
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| 摘 要： | 目的]研究黑唇鼠兔乳酸脱氢酶C（LDH-C）基因的原核表达及重组蛋白的纯化。方法]采用RT-PCR方法克隆鼠兔LDH-C基因,并将其连接到表达载体pET-32a上,构建鼠兔LDH-C基因的大肠杆菌BL21（DE3）工程菌,通过IPTG诱导表达,表达产物用SDS-PAGE进行分析,使用亲和层析方法进行蛋白纯化。结果]RT-PCR扩增出1条约1.0 kbp的条带,与预期结果相符;重组表达载体经PCR和双酶切鉴定,产生1个约1.0 kbp的目的片段,表明表达载体构建成功;表达产物经SDS-PAGE分析,得到了分子量略大于45.0kDa以包涵体形式存在的融合蛋白;通过镍亲和层析柱进行纯化,得到了较纯的融合蛋白。结论]黑唇鼠兔LDH-C基因被成功克隆和表达。 更多还原
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| 关 键 词： | 黑唇鼠兔 乳酸脱氢酶C 原核表达 蛋白纯化 |
Prokaryotic Expression of Lactate Dehydrogenase C of Black-Lipped Pika in Western Sichuan Plateau |
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| ZHANG Qing-lian et al.Prokaryotic Expression of Lactate Dehydrogenase C of Black-Lipped Pika in Western Sichuan Plateau[J].Journal of Anhui Agricultural Sciences,2011,39(14):8622-8623,8626. |
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| Authors: | ZHANG Qing-lian |
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| Institution: | ZHANG Qing-lian et al(Staff Room of Clinical Biochemistry,School of Medical Examination,Chengdu Medical College,Chengdu,Sichuan 610083) |
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| Abstract: | Objective] To express lactate dehydrogenase C(LDH-C)gene in prokaryotic system and then purify the recombinant protein.Method] The LDH-C gene was amplified from black-lipped pika(Ochotona curzoniae)by RT-PCR and ligated to the expression vector pET-32a.The E.coli BL21(DE3) carrying the LDH-C gene was obtained and induced by IPTG.The expression products were analyzed by SDS-PAGE and purified by affinity chromatography.Result] An approximately 1.0 kbp band appeared in the RT-PCR products as theoretically e... |
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| Keywords: | Black-lipped pika Lactate dehydrogenase C Prokaryotic expression Protein expression |
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