位置:成果数据库 > 期刊 > 期刊详情页
E型沙眼衣原体主要外膜蛋白多克隆抗体的制备、鉴定和初步应用
  • ISSN号:1000-6680
  • 期刊名称:《中华传染病杂志》
  • 时间:0
  • 分类:R392-33[医药卫生—免疫学;医药卫生—基础医学]
  • 作者机构:[1]天津医科大学总医院皮肤科天津性传播疾病研究所,300052
  • 相关基金:国家自然科学基金资助项目(30671879)
中文摘要:

目的表达E型沙眼衣原体主要外膜蛋白(MOMP)并制备其兔源多克隆抗体。方法诱导表达实验室构建的MOMP/pGEX6p-1重组质粒,通过胶回收进行目的蛋白的纯化。MOMP免疫新西兰家兔制备多克隆抗体,ELISA法测定抗体效价,Western印迹、细胞免疫荧光方法鉴定抗体的特异性。结果在大肠埃希菌中成功表达融合蛋白谷胱甘肽巯基转移酶(GST)-MOMP。ELISA法检测抗体的效价达到1:12800,Western印迹结果显示抗体可与原核表达的MOMP特异性结合,细胞免疫荧光检测结果显示抗体可与体外培养的沙眼衣原体特异性结合。结论成功表达了E型沙眼衣原体的MOMP,并制备了高效价、高特异性的抗MOMP抗体,为沙眼衣原体的检测和临床研究提供依据。

英文摘要:

Objective To express major outer membrane protein (MOMP) of Chlarnydia trachornatis E and to prepare rabbit polyclonal antibody. Methods The recombinant plasmid MOMP/ pGEX6p-1 prepared by our lab was introduced into E. coll. The protein was expressed and purified by gel recycling, then was injected into New Zealand rabbits to produce polyclonal antibodies. Enzyme linked immunosorbent assay (ELISA) was used to detect the titer of antibody. The antibody specificity was identified by Western blot and immunofluorescence. Results The fusion recombinant protein glutathione S-transferase (GST)-MOMP was successfully expressed in E. coli. The titer of antibody was 1:12 800 as determined by ELISA. The polyclonal antibody could specifically bind to the recombinant protein detected by Western blot and to the endogenous MOMP of Chlamydia trachoznatis in vitro detected by immunofluorescence. Conclusions The recombinant MOMP is successfully expressed and the MOMP antibody with high titer and high specificity is obtained, which will be helpful for Chlamydia trachomatis detection and related clinical research.

同期刊论文项目
同项目期刊论文
期刊信息
  • 《中华传染病杂志》
  • 中国科技核心期刊
  • 主管单位:中国科协
  • 主办单位:中华医学会
  • 主编:
  • 地址:上海市北京西路1623号
  • 邮编:200040
  • 邮箱:
  • 电话:021-62670744
  • 国际标准刊号:ISSN:1000-6680
  • 国内统一刊号:ISSN:31-1365/R
  • 邮发代号:4-352
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:23862