| 人精子蛋白FSCB的表达、纯化和多克隆抗体制备 |
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| 引用本文： | 张军,靳风烁,何畏,万江华,粱培禾,李彦锋. 人精子蛋白FSCB的表达、纯化和多克隆抗体制备[J]. 医学争鸣, 2009, 0(21): 2271-2274 |
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| 作者姓名： | 张军 靳风烁 何畏 万江华 粱培禾 李彦锋 |
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| 作者单位： | 第三军医大学大坪医院野战外科研究所泌尿外科;第三军医大学西南医院妇产科; |
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| 基金项目： | 重庆市自然科学基金(CSTC,2007BB5081) |
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| 摘 要： | 目的：构建表达重组人精子蛋白FSCB的重组质粒,获得其纯化重组蛋白并制备其多克隆抗体.方法：分别成功构建了表达人精子全长FSCB蛋白和截短FSCB（FSCBt）蛋白的重组菌株BL21,IPTG诱导表达,镍柱亲和层析法和分子筛层析法纯化目的蛋白FSCBt,免疫接种雌性家兔制备多克隆抗体.结果：重组菌成功诱导表达了重组蛋白FSCB和FSCBt,相对表观分子量分别为230000和30000,重组蛋白FSCBt经纯化后纯度达到95%,制备的多克隆抗体效价达到1∶105,Western Blot显示多克隆抗体与重组蛋白FSCB和FSCBt均具有良好的反应性.结论：人精子蛋白FSCB在重组菌株中得到表达,成功制备高效价的多克隆抗体,为进一步研究该蛋白在人体组织中的表达、分布以及生物学功能奠定了实验基础.
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| 关 键 词： | FSCB 多克隆抗体 蛋白质纯化 基因重组 |
Expression,purification and polyclonal antibody preparation of human sperm protein FSCB |
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| ZHANG Jun,JIN Feng-Shuo,HE Wei,WAN Jiang-Hua,LIANG Pei-He,LI Yan-Feng. Expression,purification and polyclonal antibody preparation of human sperm protein FSCB[J]. Negative, 2009, 0(21): 2271-2274 |
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| Authors: | ZHANG Jun JIN Feng-Shuo HE Wei WAN Jiang-Hua LIANG Pei-He LI Yan-Feng |
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| Affiliation: | ZHANG Jun1,JIN Feng-Shuo1,HE Wei2,WAN Jiang-Hua1,LIANG Pei-He1,LI Yan-Feng11Department of Urology,Institute of Field Surgery,Daping Hospital,Chongqing 400042,2Department of Gynaecology and Obstetrics,Southwest Hospital,Third Military Medical University,Chongqing 400038,China |
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| Abstract: | AIM:To express the recombinant protein and prepare the polyclonal antibody(PAb) of human sperm protein FSCB. METHODS:Two recombinant bacterium BL21 stains expressing human full length FSCB and truncated FSCB (FSCBt) proteins were successfully constructed,and the FSCBt protein was purified by Ni2+ affinity chromatography and molecular sieve chromatography. The specific PAb of FSCB was prepared by immunizing female rabbits. RESULTS:The recombinant protein FSCB and FSCBt could be expressed stably by the recomb... |
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| Keywords: | FSCB polyclonal antibody protein purification gene recombination |
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