Hsp70和汉滩病毒S基因融合蛋白的原核表达及免疫小鼠的初步研究.pdfVIP

21 1 21 111-15 2006 1 VIROLOGICA SINICA January 2006 Hsp70 S * 1 2 1** 3 1 1. 710032; 2. 710032; 3. 710032 Prokaryotic Expression and Purification of Hsp70-NP and Its Immunity 1 2 1** 3 1 LI Jing YE Zheng-xu YANG Shou-jing GAO Juan LIU Yan-fang 1.Department of Pathology, Xian 710032, China; 2. Department of Orthopaedics, Xian 710032, China; 3. Department of Gastroenterology, Xi Jing Hospital, Fourth Military Medical University, Xian 710032, China Abstract Recombinant fusion prokaryotic expression vector pGEX-4T-1/hsp70-S was constructed by cloning genes Hsp70 and Hantaan virus S gene coding region into pGEX-4T and was expressed in E. coli. The Hsp70-NP fusion protein was separated and purified with GSTrapFF purification system. We also constructed the prokaryotic expression vectors pGEX-4T-1/hsp70 and pET28a/S to prepare the purified protein Hsp70 and NP. Then BANB/c mice were vaccinated with the protein Hsp70-NP, Hsp70 and NP separately. ELISA results showed that both Hsp70-Npand NP could induce specific antibodies against NP. The specific antibody titers stimulated by Hsp70-NP were obviously higher than that of NP. The lymphoproliferative responses of spleen cells clearly showed that the spleen cells from both groups of mice were able to proliferate in the presence of NP protein. The stimulation indexes of splenocytes to NP induced by Hsp70-NP were significantly higher than that induced by NP. It indicated that linkage of Hsp70 to NP enhanced the immune response to Hantaan virus nucleocapsid protein. Key words Heat shock protein; Hantaan Virus S gene; Prokaryotic expre