摘要
目的构建含有CD24基因的重组质粒,在人胚肾上皮细胞Hek293中表达出具有免疫原性的抗原。方法用PCR方法扩增CD24基因,克隆到载体pYD5-hFC中,获得重组质粒pYD5-CD24,转化DH10α感受态细胞中,挑选克隆抽取质粒,以脂质体介导将质粒转染Hek293细胞,72 h后收集细胞培养上清,Western-Blot分析鉴定。结果构建了含有CD24基因的重组质粒,并在人胚肾上皮细胞Hek293中获得带有hFC标签的CD24蛋白。结论本研究成功表达出了具有免疫原性的CD24蛋白。
Objective To construct the recombinant plasmid with gene CD24 and to express immunogenic protein in human embryo kidney Hek293 cells. Methods CD24 gene was cloned into the vector pYD5-hFC by PRC method. The required recombinant plasmids pYD5-CD24 transformed into the GH10α component cells. Then the selected recombinant plasmids were transfected with lipofectamine reagent into Hek293 cells. After 72 hours,the protein in the culture supernatant of the Hek293 cells were collected and analyzed by western blot. Results The recombinant plasmid transfected with CD24 gene were constructed. It was detected out that the CD24 protein with hFC sign were expressed in human embryo kidney Hek293 cells. Conclusion The CD24 gene expressed successfully in the transfected Hek293 cells and had good immunogenic activity.
出处
《中国现代医生》
2012年第29期47-48,51,共3页
China Modern Doctor
基金
国家自然科学基金(81072008)
