CAJ | 학술논문

以马耳他布鲁菌基因组为模板,设计引物,扩增17KD膜蛋白基因全长,获得422bp长的片段。将该片段直接克隆于原核表达载体PGEX-4T-2,成功构建阳性重组表达载体。IPTG诱导表达,经SDS-PAGE和Western blot检测,在大肠杆菌中成功表达大小为43KD的蛋白,并且该蛋白与布鲁菌病临床阳性动物血清产生特异性结合反应。通过17KD蛋白的成功表达,试图评价该蛋白在布鲁菌病诊断用抗原及基因工程疫苗研制过程中应用的可行性。
이마이타포로균기인조위모판,설계인물,확증17KD막단백기인전장,획득422bp장적편단。장해편단직접극륭우원핵표체재체PGEX-4T-2,성공구건양성중조표체재체。IPTG유도표체,경SDS-PAGE화Western blot검측,재대장간균중성공표체대소위43KD적단백,병차해단백여포로균병림상양성동물혈청산생특이성결합반응。통과17KD단백적성공표체,시도평개해단백재포로균병진단용항원급기인공정역묘연제과정중응용적가행성。
Primers were devised according to protein 17KD gene sequence,then took 16M brucella as template,amplified the gene fragment by PCR.Directly cloned the fragment into the prokaryotic expression vector pGEX-4T-2,obtained a recombinant positive plasmid.Recombinant 17KD protein was successfully expressed in E.coli.and reacted with clinical positive serum of brucellosis.With the recombinant 17KD protein,try to evaluate the role of the recombinant protein in the brucellosis diagnotic and gene vaccine.