【摘要】 目的:构建CD28分子胞外区噬菌体展示载体,并对其展示的CD28抗原性进行检测。方法:采用RT-PCR方法扩增CD28胞外区,将其重组于噬菌粒载体pComb3HSS,构建的重组载体与辅助噬菌体VCSM13共转染受体菌,使CD28胞外区展示在噬菌体表面。用ELISA和流式细胞仪检测展示在噬菌体上CD28的抗原性。结果:成功构建了CD28的重组噬菌体展示载体pComb3HSS-CD28。噬菌体展示的CD28可与抗CD28抗体结合,也可与JurkatT细胞表面的CD28竞争性结合抗CD28的抗体。结论:噬菌体展示的CD28具有抗原性。更多还原

【Abstract】 Objective: To construct a phage display vecetor for human CD28 and display CD28 on phage. Methods: CD28 extra-cellular gene was amplified from Jurkat cells by RT-PCR technique and cloned into pComb3HSS. The recombinant pComb3H-CD28 was co-infected into bacteria with help phage VCSM13 and CD28 extra-cellular domain homodimer was displayed on the phage surface. The antigenicity of CD28 displayed on phage was detected by ELISA and FCM. Results: CD28 RT-PCR products were 420 bp, the recombined plasmid was named pComb3H-CD28, and CD28 homodimer was displayed on phage. ELISA and FCM detection showed that phage displayed CD28 homodimer possessed antigenicity of human CD28. Conclusion: The studies demonstrate that phage display system can display human CD28 extra-cellular domain homodimer with antigenicity.更多还原