| 山羊PTHrP基因的多克隆抗体制备和组织表达图谱分析 |
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| 引用本文： | 郑惠玲,杨振宇,祝珍珍,安俊辉,邢瑞芳.山羊PTHrP基因的多克隆抗体制备和组织表达图谱分析[J].畜牧兽医学报,2011,42(2). |
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| 作者姓名： | 郑惠玲 杨振宇 祝珍珍 安俊辉 邢瑞芳 |
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| 作者单位： | 西北农林科技大学动物科技学院,陕西省农业分子生物学重点实验室,杨凌,712100 |
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| 基金项目： | 转基因生物新品种培育重大专项(2009ZX08009-162B); 中央高校基本科研业务费专项资金(Z109021002)资助 |
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| 摘 要： | 旨在获得山羊PTHrP多克隆抗体,检测其在山羊各组织中的表达图谱,以便进一步研究山羊PTHrP的功能。本研究构建山羊PTHrP基因的原核表达载体,转化E.coliBL21(DE3)菌株,IPTG诱导表达获得融合蛋白,SDS-PAGE电泳检测融合蛋白,然后将融合蛋白纯化后多点免疫大耳白兔,制备多克隆抗体,最后使用West-ern blot检测PTHrP在山羊各组织中的表达。结果,酶切和测序显示原核表达载体构建成功;SDS-PAGE电泳结果说明融合蛋白(约36.5 ku)被成功纯化;间接ELISA检测所制备的抗体效价达1∶51 200;表达谱显示PTHrP在山羊乳腺、肾脏、垂体、脑、心脏、肝脏、骨、肌肉、脾脏等组织中均有表达。本研究成功制备了山羊PTHrP的多克隆抗体,发现PTHrP在山羊各组织中广泛表达。
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| 关 键 词： | 甲状旁腺相关肽 原核表达 多克隆抗体 组织表达图谱 山羊 |
Preparation of Goat PTHrP Polyclonal Antibody and Analysis of PTHrP Expression Map in Tissues |
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| ZHENG Hui-ling,YANG Zhen-yu,ZHU Zhen-zhen,AN Jun-hui,XING Rui-fang.Preparation of Goat PTHrP Polyclonal Antibody and Analysis of PTHrP Expression Map in Tissues[J].Acta Veterinaria et Zootechnica Sinica,2011,42(2). |
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| Authors: | ZHENG Hui-ling YANG Zhen-yu ZHU Zhen-zhen AN Jun-hui XING Rui-fang |
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| Institution: | ZHENG Hui-ling,YANG Zhen-yu,ZHU Zhen-zhen,AN Jun-hui,XING Rui-fang (Key Laboratory for Molecule Biology of Agriculture,College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China) |
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| Abstract: | To further understand the biological function of goat PTHrP gene,the PTHrP polyclonal antibody was prepared and the tissue expression profile of PTHrP was analyzed in this study.Prokaryotic expression vector was constructed and expressed in E.coli BL21(DE3) host cells.Fusion protein was purified and used to immunize rabbit.Western blot was performed to assay the tissue expression profile.Restriction enzyme mapping and sequencing showed that prokaryotic expression vector was constructed successfully.SDS-PAGE... |
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| Keywords: | parathyroid hormone related protein prokaryotic expression polyclonal antibody tissue specific expression profile goat |
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