| 猪MyD88基因的原核表达 |
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| 引用本文： | 曾爽,陈国华,崔青,房永祥,景志忠.猪MyD88基因的原核表达[J].动物医学进展,2010,31(12). |
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| 作者姓名： | 曾爽 陈国华 崔青 房永祥 景志忠 |
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| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃兰州,730070;中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,农业部兽医公共卫生重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州,730046
2. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,农业部兽医公共卫生重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州,730046 |
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| 基金项目： | 国家自然科学基金项目(308718840); 国家高新技术研究发展计划(863)项目(2006AA10A203) |
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| 摘 要： | 采用PCR方法从pGEM-pMyD88重组质粒中克隆了猪MyD88分子全长基因,构建了无信号肽序列的原核表达载体pET-30a-pMyD88-ED,在不同IPTG浓度和时间下进行诱导表达,利用切胶洗脱法纯化表达的融合蛋白。结果表明,在IPTG诱导浓度为0.05 mmol/L,诱导时间为6 h时表达量达到最大,SDS-PAGE分析表明表达出约35 ku的融合蛋白,主要以包涵体的形式存在;通过切胶纯化后,融合蛋白的纯度可达90%;Western blot分析显示,表达的融合蛋白能被抗His标签的单克隆抗体识别,说明目的蛋白在大肠埃希菌中得到了成功表达。
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| 关 键 词： | 猪MyD88分子 原核表达 大肠埃希菌 |
Prokaryotic Expression of Porcine Mydoid Differentiation Factor 88 |
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| ZENG Shuang,CHEN Guo-hua,CUI Qing,FANG Yong-xiang,JING Zhi-zhong.Prokaryotic Expression of Porcine Mydoid Differentiation Factor 88[J].Progress In Veterinary Medicine,2010,31(12). |
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| Authors: | ZENG Shuang CHEN Guo-hua CUI Qing FANG Yong-xiang JING Zhi-zhong |
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| Institution: | ZENG Shuang1,2,CHEN Guo-hua2,CUI Qing1,FANG Yong-xiang2,JING Zhi-zhong1,2(1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou,Gansu,730070,China,2.Key Laboratory of Veterinary Parasitology of Gansu Province,Key Laboratory of Veterinary Public Health of Ministry of Agriculture,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,CAAS,730046,China) |
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| Abstract: | Porcine MyD88 molecule fragment(pMyD88-ED)was amplied by PCR method from the pGEM-T-pMyD88 recombinant plasmid,the pMyD88-ED was cloned into pET-30a vector and expressed in Escherichia coli.The recombinant strain was induced in different IPTG concentrations and different time.The results showed that the optimum concentrations of IPTG is 0.05 mmol/L,the optimum time is 6 h.SDS-PAGE analysis showed that the recombinant protein was expressed with a molecular weight of 35 ku and existed mainly in the form of in... |
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| Keywords: | porcine mydoid differentiation factor 88(pMyD88) prokaryotic expression Escherichia coli |
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