Aim: To design and construct an HIV-1 subunit vaccine containing the N-terminal heptad repeat (NHR) domain N51 in gp41 of the HIV-1 CRF01_AE recombinant subtype in China and study its immunogenicity.
Methods: Two pairs of primers were designed to amplify DNA fragment encoding N51Fd gene, which was then subcloned into pFUSE-hIgG1-Fc2 vector. The recombinant plasmid pFUSE/N51Fd was confirmed by DNA sequencing. Western blotting was used to measure the correct expression of the recombinant protein N51FdFc-AE. The N51FdFc-AE protein was used to immunize BALB/c mice, and the specific antibody response was measured by ELISA.
Results: A recombinant plasmid pFUSE/N51Fd was successfully constructed and the N51FdFc-AE recombinant protein was expressed effectively in 293T cells. The purified N51FdFc-AE recombinant protein (35 kD) was detected by Western blotting with rabbit anti-gp41 N/C peptide antibodies. The mouse anti-sera could specifically recognize the antigens derived from gp41 NHR. The titers of the specific antibodies were up to 1:102 400 with an average titer of 1:51 200.
Conclusion: The recombinant N51FdFc-AE protein can effectively induce the gp41 NHR-specific immune responses in BALB/c mice and could be used as an immunogen for design of HIV-1 subunit vaccine.
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