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| 利用免疫蛋白质组学鉴定炭疽杆菌蛋白MetK及其免疫原性验证 | |
| 引用本文： | 童超,刘先凯,任静晓,王东澍,高志奇,冯尔玲,朱力,廖祥儒,王恒樑.利用免疫蛋白质组学鉴定炭疽杆菌蛋白MetK及其免疫原性验证[J].军事医学科学院院刊,2012,36(2):115-119. |
| 作者姓名： | 童超 刘先凯 任静晓 王东澍 高志奇 冯尔玲 朱力 廖祥儒 王恒樑 |
| 作者单位： | 1. 江南大学工业生物技术教育部重点实验室,江苏无锡,214122;军事医学科学院生物工程研究所,病原微生物生物安全国家重点实验室,北京,100071 2. 军事医学科学院生物工程研究所,病原微生物生物安全国家重点实验室,北京,100071 3. 江南大学工业生物技术教育部重点实验室,江苏无锡,214122 |
| 摘 要： | 目的验证免疫蛋白质组中鉴定到的S-腺苷甲硫氨酸MetK蛋白的免疫原性。方法免疫蛋白质组学的方法鉴定到MetK蛋白,通过PCR方法扩增metK基因全长,然后将其克隆到原核表达载体pET32a中,转化大肠杆菌DH5α,测序正确后提取质粒转入BL21（DE3）中进行诱导表达。再通过Ni-NTA柱纯化MetK蛋白,纯化后的蛋白免疫SPF级的BALB/c小鼠,制备多克隆抗体,通过多克隆抗体与炭疽芽孢杆菌A16R全菌蛋白Western印迹检测,验证MetK蛋白的免疫原性。结果与结论成功表达了MetK蛋白,制备了其多克隆抗体,为免疫蛋白质组学鉴定到的蛋白点进行验证,也为研究MetK蛋白的生物学功能研究提供了条件。 |
| 关 键 词： | 免疫蛋白质组学 metK 芽孢杆菌 炭疽 基因表达 多克隆抗体 |
Immunogenic protein MetK of Bacillus anthracis identified by immunoproteomics |
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| TONG Chao , LIU Xian-kai , REN Jing-xiao , WANG Dong-shu , GAO Zhi-qi , FENG Er-ling , ZHU Li , LIAO Xiang-ru , WANG Heng-liang.Immunogenic protein MetK of Bacillus anthracis identified by immunoproteomics[J].Bulletin of the Academy of Military Medical Sciences,2012,36(2):115-119. | |
| Authors: | TONG Chao LIU Xian-kai REN Jing-xiao WANG Dong-shu GAO Zhi-qi FENG Er-ling ZHU Li LIAO Xiang-ru WANG Heng-liang |
| Institution: | liang (1. Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China ;2. State Key Laborary of Pathogen and B iosecurity, Beijing Institute of Biotechnology, Beijing 100071, China) |
| Abstract: | Objective To confirm the immunogenicity of Bacillus anthracis MetK identified by immunoproteomics study. Methods Immunoproteomics study of B. anthracis A16R revealed a protein MetK. metK genes were amplified by PCR and cloned into prokaryotic expression vector pET32a. Recombinant plasmid pET32a-metK was transformed into E. coli BL21 （DE3）, MetK was expressed by inducing IPTG and purified by Ni affinity chromatography. Polyclonal antibody in mice was prepared by intraperitoneal injection. Western blot was used to assay the immunogenicity of MetK protein. Results and Conclusion MetK protein was successfully expressed and polyclonal antibody was produced, The polyclonal antisera can be used to study the function of MetK protein. |
| Keywords: | immunoproteomics rnetK Bacillus anthracis gene expression polyclonal antibody |
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