| 大鼠Fcε3-4基因克隆及原核表达 |
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| 引用本文： | 张艳芬,时海浪,王培莹,乔晓强,刘中成.大鼠Fcε3-4基因克隆及原核表达[J].河北大学学报(自然科学版),2012,32(4):387-392. |
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| 作者姓名： | 张艳芬 时海浪 王培莹 乔晓强 刘中成 |
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| 作者单位： | 1. 河北大学实验室管理办公室,河北保定,071002
2. 河北大学药学院,河北保定,071002 |
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| 基金项目： | 河北省自然科学基金资助项目(C2010000248);河北省科学技术研究与发展计划项目(11275530);河北大学大学生科技创新项目(2011089) |
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| 摘 要： | 为了获得重组Fcε3-4蛋白,从变态反应性哮喘大鼠脾组织中提取总RNA,通过RT-PCR方法克隆大鼠Fcε3-4基因,构建成原核表达载体pET17b-Fcε3-4,并转化大肠杆菌进行诱导表达,Fcε3-4蛋白主要以包涵体形式存在.经Ni-NAT亲和层析柱对尿素溶解的Fcε3-4蛋白进行了纯化,并利用降低尿素梯度的方法柱上对纯化蛋白进行了复性.经Western Blotting和ELISA鉴定,Fcε3-4蛋白具有Fcε的免疫特异性,能被抗大鼠Fcε抗体特异性识别,为下一步研究该蛋白奠定了基础.
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| 关 键 词： | 大鼠 Fcε3-4基因 克隆 表达 免疫特异性 |
Cloning and prokaryotic expression of rat Fcε3-4 gene |
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| ZHANG Yan-fen
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SHI Hai-lang
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WANG Pei-ying
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QIAO Xiao-qiang
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LIU Zhong-cheng.Cloning and prokaryotic expression of rat Fcε3-4 gene[J].Journal of Hebei University (Natural Science Edition),2012,32(4):387-392. |
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| Authors: | ZHANG Yan-fen SHI Hai-lang WANG Pei-ying QIAO Xiao-qiang LIU Zhong-cheng |
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| Institution: | 1.Laboratory Management Office,Hebei University,Baoding 071002,China; 2.College of Pharmaceutical Science,Hebei University,Baoding 071002,China) |
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| Abstract: | In order to obtain recombinant Fcε3-4 protein the Fcε3-4 gene was amplified by the method of RT-PCR from the total RNA extracted from a fresh spleen of allergic asthma rat,then cloned into expression vector pET-17b.The Fcε3-4 protein was expressed as inclusion body in E.coli BL21.The denatured Fcε3-4 protein was dissolved in 8 mol/L urea,then purified and refolded in lower urea gradient method on Ni-NAT column.The expressed Fcε3-4 owned the epitope recognised by anti-Fcε antibody through Western Blotting and ELISA analysis,which would be a basis for further studies of the Fcε3-4 protein. |
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| Keywords: | rat Fcε3-4 gene clone expression immunologic specificity |
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