CAJ | 학술논문

根据GenBank SS2(Streptococcus suis serotype 2,SS2)epf基因序列设计引物,克隆epf基因片段并进行序列分析,同时亚克隆到GST融合蛋白表达载体pGEX4T-2上,将构建的原核表达质粒pGEx4T-2-epf导入大肠杆菌(Eacterium coli)TG1中,诱导融合蛋白EF-GST的表达.经亲和层析、凝血酶切割后,获得纯化的EF为抗原蛋白,用其免疫家兔制备抗血清,成功制备了EF蛋白及其特异的多克隆抗体,为进一步研究EF的功能提供了有用的工具.
근거GenBank SS2(Streptococcus suis serotype 2,SS2)epf기인서렬설계인물,극륭epf기인편단병진행서렬분석,동시아극륭도GST융합단백표체재체pGEX4T-2상,장구건적원핵표체질립pGEx4T-2-epf도입대장간균(Eacterium coli)TG1중,유도융합단백EF-GST적표체.경친화층석、응혈매절할후,획득순화적EF위항원단백,용기면역가토제비항혈청,성공제비료EF단백급기특이적다극륭항체,위진일보연구EF적공능제공료유용적공구.
The epf gene fragment was amplified by PCR from and cloned into prokaryotic expression plasmid pGEX4T-2 to form pGEX4T-2-epf. The expression of recombinant protein with GST was induced in Eacteriwn coli TGI and the fusion protein (EF-GST) was purified by affinity chromatography,and then GST was cut with thrombin protease. The ployclonal anti-EF antibody was prepared by immunizing rabbits with the purified protein,expressed and purified proteins were analyzed by SDS-PAGE and confirmed by Western blotting. These results suggested that the purified EF protein and the poly-clonal antibody could be used for further studies of SS2 (Streptococcus suis serotype 2).