IL-32原核表达载体的构建及表达
Construction of the Prokaryotic Expression Vector of IL-32 Gene and Its Expression
摘要
目的构建人IL-32原核表达载体,并进行初步诱导表达。方法PCR扩增IL-32基因并将其与原核表达质粒pET32a(+)连接,重组pET32a—IL-32经PCR、酶切及测序鉴定后转入表达菌株BL21(DE3)进行融合蛋白的少量诱导表达,SDS—PAGE和Westem Blotting检测目的蛋白表达及水平。结果含有重组质粒的表达菌株经IPTG诱导后获得高效表达,融合蛋白主要存在于包涵体中,重组蛋白表达水平占总蛋白的27.5%;Western Blotting分析表明该蛋白可与特异性抗体发生结合。结论构建了人IL-32基因的原核表达载体,获得IL-32融合蛋白。
出处
《潍坊医学院学报》
2010年第5期321-323,共3页
Acta Academiae Medicinae Weifang
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