摘要
目的:利用原核表达系统表达BLCAP融合蛋白并制备其多克隆抗体。方法:构建原核表达重组质粒pET32a(+)-BLCAP并转化到宿主菌BL21中,通过IPTG诱导表达并采用亲和层析的方法纯化和SDS-PAGE鉴定后免疫日本大耳白兔,制备BLCAP蛋白多克隆抗体并检测其特异性。结果:经过IPTG诱导,获得分子量大小约为28ku的融合蛋白,用纯化得到的融合蛋白免疫日本大耳白兔后得到抗BLCAP蛋白的多克隆抗体,通过Western blot检测证明该抗体能够与BLCAP融合蛋白发生特异性结合。结论:重组质粒表达的BLCAP融合蛋白具有良好的抗原性,制备的抗BLCAP的多克隆抗体特异性较好,为今后进一步研究BLCAP蛋白性质与功能奠定了基础。
Objective:To express BLCAP fusion protein in E.coli BL21(DE3) and prepare polyclonal antibody. Methods:The prokaryotic expression plasmid pET32a(+)-BLCAP was constructed and the positive recombinant plasmid was transformed into BL21(DE3) and induced by IPTG. After the soluble protein was purified by affinity chromatography and identified by SDS-PAGE,the rabbits were immunized with the fusion protein and the antiserum was obtained. The specificity of the rabbit’s antiserum was measured by Western bloting. Re...
关键词
BLCAP
宫颈癌
原核表达
多克隆抗体
BLCAP
Cervical carcinoma
Prokaryotic expression
Polyclonal antibody
