Citation: LI Jing, YE Zheng-xu, YANG Shou-jing, GAO Juan, LIU Yan-fang. Prokaryotic Expression and Purification of Hsp70-NP and Its Immunity .VIROLOGICA SINICA, 2006, 21(1) : 11-15.

Prokaryotic Expression and Purification of Hsp70-NP and Its Immunity

  • Corresponding author: YANG Shou-jing, 
  • Available online: 20 January 2006
  • 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.

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    Prokaryotic Expression and Purification of Hsp70-NP and Its Immunity

      Corresponding author: YANG Shou-jing,
    • 1. Department of Pathology, 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.

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