| 人脂联素原核表达纯化及活性检测 |
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| 引用本文： | 王云龙,李永超,昌静峰,李玉林,王国强,邓黎黎,陈冬焕,董彩文,孙新城,刘旺根,梁晓艳.人脂联素原核表达纯化及活性检测[J].动物医学进展,2011(10):36-39. |
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| 作者姓名： | 王云龙 李永超 昌静峰 李玉林 王国强 邓黎黎 陈冬焕 董彩文 孙新城 刘旺根 梁晓艳 |
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| 作者单位： | 河南工业大学;郑州职业技术学院;河南省生物工程技术研究中心; |
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| 基金项目： | 河南省科技创新团队和郑州市科技创新团队资助项目 |
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| 摘 要： | 为了克隆人脂联素基因(ACRP30),获得有免疫活性的重组人脂联素,以人皮下脂肪组织为材料,提取总RNA,通过RT-PCR得到ACRP30基因.构建重组质粒pET-CKS-ACRP30,转化表达茵BL21(DE3),以IPTG诱导表达人脂联素重组蛋白,利用镍亲和层析纯化,Western blot鉴定其活性,双抗体夹心法...
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| 关 键 词： | 人脂联素 原核表达 免疫活性 |
Prokaryotic Expression and Purification of Human Adiponectin and Its Immunological Activity Detection |
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| WANG Yun-long,LI Yong-chao,CHANG Jing-feng,LI Yu-lin,WANG Guo-qiang,DENG Li-li,CHEN Dong-huan,DONG Cai-wen,SUN Xin-cheng,LIU Wang-gen,LIANG Xiao-yan.Prokaryotic Expression and Purification of Human Adiponectin and Its Immunological Activity Detection[J].Progress In Veterinary Medicine,2011(10):36-39. |
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| Authors: | WANG Yun-long LI Yong-chao CHANG Jing-feng LI Yu-lin WANG Guo-qiang DENG Li-li CHEN Dong-huan DONG Cai-wen SUN Xin-cheng LIU Wang-gen LIANG Xiao-yan |
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| Institution: | WANG Yun-long1,2,LI Yong-chao1,CHANG Jing-feng3,LI Yu-lin3,WANG Guo-qiang3,DENG Li-li3,CHEN Dong-huan3,DONG Cai-wen3,SUN Xin-cheng3,LIU Wang-gen3,LIANG Xiao-yan3(1.Henan University of Technology,Zhengzhou,Henan,450001,2.Zhengzhou Technical College,450121,China,3.Henan Biotechnology Research Center,China) |
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| Abstract: | To clone the human adiponectin(ACRP30) cDNA and obtain the recombinant adiponectin with immunological activity,fresh human fat tissue was used for extracting total RNA and amplificating the adiponectin cDNA through RT-PCR.The recombinant plasmid pET-CKS-ACRP30 was constructed and confirmed,and it was transduced into BL21(DE3).Then the transformation was used for induced expression.The recombinant protein was identified by Western blotting and purified by Ni affinity chromatography.Finally,the sandwich ELISA... |
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| Keywords: | human adiponectin prokaryotic expression immunological activity |
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