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| 山羊痘病毒p32基因的克隆、表达及单克隆抗体制备 | |
| 引用本文： | 王永志,苏颖,米丽娟,付云红,范志强,扈荣良,赵玉民.山羊痘病毒p32基因的克隆、表达及单克隆抗体制备[J].中国畜牧兽医,2011,38(9):101-105. |
| 作者姓名： | 王永志 苏颖 米丽娟 付云红 范志强 扈荣良 赵玉民 |
| 作者单位： | 1. 吉林省农业科学院,吉林长春130124;中国人民解放军军事医学科学院军事兽医研究所,吉林长春 130062 2. 吉林省农业科学院,吉林长春,130124 3. 吉林农业大学动物科学技术学院,吉林长春,130118 4. 中国人民解放军军事医学科学院军事兽医研究所,吉林长春,130062 |
| 基金项目： | 吉林省农业科学院博士科研启动基金(0100434) |
| 摘 要： | 本研究旨在表达山羊痘病毒p32蛋白、制备p32蛋白单克隆抗体。通过PCR克隆p32基因,连接到pMD18-T并测序;酶切、连接后将p32基因克隆到pET28a(+)中,将重组质粒转化大肠杆菌Rosseta,在IPTG诱导下成功表达p32蛋白;利用亲和层析法纯化p32蛋白,用纯化蛋白免疫BALB/c小鼠后制备p32蛋白单克隆抗体,获得22株识别重组蛋白的单抗,通过间接免疫荧光鉴定,其中3株能识别病毒的p32蛋白。山羊痘病毒p32蛋白的表达和单克隆抗体的制备为山羊痘病毒抗原、抗体检测提供了生物材料。 |
| 关 键 词： | 山羊痘病毒 p32基因 单克隆抗体 |
| 收稿时间： | 2011-01-21 |
Clone,Expression and Monoclonal Antibody Preparation of p32 Gene of GPV |
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| WANG Yong-zhi,SU Ying,MI Li-juan,FU Yun-hong,FAN Zhi-qiang,HU Rong-liang,ZHAO Yu-min.Clone,Expression and Monoclonal Antibody Preparation of p32 Gene of GPV[J].China Animal Husbandry & Veterinary Medicine,2011,38(9):101-105. | |
| Authors: | WANG Yong-zhi SU Ying MI Li-juan FU Yun-hong FAN Zhi-qiang HU Rong-liang ZHAO Yu-min |
| Institution: | 1. Jilin Academy of Agricultural Sciences, Changchun 130124, China;2. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China;3. Veterinary Institute, Academy of Military Medical Sciences of PLA, Changchun 130062, China |
| Abstract: | For expressing and preparing monoclonal antibody of GPV p32 protein,p32 gene was cloned and inserted into plasmid pET28a(+),and expressed efficiently in E.coli Rosseta by IPTG induction.After purification by affinity chromatograph,p32 protein was immunized as antigen to BALB/c mouse.22 monoclonal antibodies which recognize recombinant p32 protein were obtained,among them,3 could recognize GPV.These monoclonal antibodies provided biomaterials for sandwich ELISA testing GPV and competitive ELISA testing GPV a... |
| Keywords: | GPV p32 gene monoclonal antibody |
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