CAJ | 학술논문

为获得狂犬病毒G基因并在大肠杆菌中进行表达，进而初步建立用于评价狂犬疫苗免疫效果的方法，根据狂犬病病毒RV（rabies virus）ERA株G基因序列设计引物，从病毒中提取出RNA，经逆转录并扩增得到RVG基因；将该基因片段定向克隆到原核表达载体pET28a（＋）中，构建重组质粒PET-RVG并转化到大肠杆菌BL21（DE3）gold中，经IPTG诱导表达并确定表达的最佳条件；Ni亲和层析柱纯化获得重组G蛋白，并以G蛋白为抗原建立检测狂犬病毒抗体的间接ELISA方法。检测结果表明，经灰度分析纯化后纯度可达96％。
위획득광견병독G기인병재대장간균중진행표체，진이초보건립용우평개광견역묘면역효과적방법，근거광견병병독RV（rabies virus）ERA주G기인서렬설계인물，종병독중제취출RNA，경역전록병확증득도RVG기인；장해기인편단정향극륭도원핵표체재체pET28a（＋）중，구건중조질립PET-RVG병전화도대장간균BL21（DE3）gold중，경IPTG유도표체병학정표체적최가조건；Ni친화층석주순화획득중조G단백，병이G단백위항원건립검측광견병독항체적간접ELISA방법。검측결과표명，경회도분석순화후순도가체96％。
In order to acquire rabies virus G gene and express it in E.coli and establish the evaluation method for rabies virus immune,according to Rabies virus ERA G gene sequence to design primer se-quences,RNA was extracted from rabies virus,the first strand cDNA was synthesized by reverse transcrip-tion.G gene fragments were amplified by PCR.Then the gene fragment was cloned to the prokaryotic ex-pression vector pET28a (+)to construct PET-RVG.The positive recombinant plasmids were transfected into E.coli BL21(DE3)gold,with IPTG induced expression and determined the best expression condition. Using Niaffinity chromatography purification with G protein,and the indirect ELISA assay for the detection of rabies virus antibodies in serum was established based on the G protein.The results showed that the puri-ty degree was 96%.