利用DNA免疫技术制备抗登革2型病毒NS2B蛋白多克隆抗体  

Preparation of polyclonal antibodies against NS2B of dengue virus serotype 2 by immunization with DNA

作　　者：刘丽梅[1] 陈宗涛[2] 高娜[3] 田衍平[3] 陈炜[2] 张俊磊[2] 王嘉丽[2] 安静[4] 

机构地区：[1]第三军医大学西南医院病理学研究所,重庆400038 [2]第三军医大学基础医学部微生物学教研室、重庆市微生物工程实验室,重庆400038 [3]第三军医大学基础医学部组织胚胎学教研室、重庆市神经科学研究所,重庆400038 [4]首都医科大学基础医学院微生物学教研室,北京100069

出　　处：《第三军医大学学报》2010年第4期327-330,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30671853,30872227)~~

摘　　要：目的构建登革2型病毒(dengue virus serotype2,DV2)非结构蛋白NS2B的真核表达质粒pCAGGS-P7/NS2B,进行DNA免疫,制备小鼠抗NS2B特异性多克隆抗体。方法PCR扩增DV2-NS2B基因片段,构建真核表达载体pCAGGS-P7/NS2B,采用脂质体介导的方法转染Vero细胞,检测目标蛋白的表达。用该表达质粒免疫BALB/c小鼠,制备抗NS2B抗血清。结果成功构建真核表达重组质粒pCAGGS-P7/NS2B,该重组质粒具有良好的免疫原性,免疫小鼠后所获的抗血清,抗体效价达1∶3200,Western blot和间接免疫荧光证实抗血清能特异性地识别DV2-NS2B蛋白。结论通过DNA免疫所制备小鼠抗DV2-NS2B抗血清能特异性地识别DV2-NS2B蛋白。Objective To construct the eukaryotic expression plasmid pCAGGS-P7/NS2B of dengue virus serotype 2 （DV2）NS2B and to prepare mouse anti-NS2B specific polyclonal antibody by immunizing mice with DNA. Methods An eukaryotic expression vector pCAGGS-P7/NS2B was constructed by PCR amplification of the NS2B gene fragment. Vero cells were transfected with lipid-mediated gene. The expression of target protein DV2 NS2B was detected in Vero cells. BALB/c mice were immunized with pCAGGS-NS2B to prepare anti-serum against NS2. Results The pCAGGS-P7/NS2B was constructed with a rather good immunogenicity. The antibody valence of anti-serum obtained from the immunized mice was 1： 3200. Western blotting and indirect immunofluorescence showed that the anti-serum could specifically recognize the DV2-NS2B protein. Conclusion Anti-serum against DV2-NS2 prepared by immunizing mice with DNA can specifically recognize the DV2-NS2B protein.

关 键 词：登革热病毒 病毒非结构蛋白质类 NS2B 小鼠 近交BALBC 

分 类 号：R392-33[医药卫生—免疫学]