title: [Characterization of the HIV-1 subunit vaccine containing the gp41 NHR domain of the HIV-1 CRF01_AE recombinant subtype in China].
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China. jpshaosl@163.com
authors.forename: Ji-ping
authors.initials: JP
authors.lastname: Shao
authors.forename: Shi-bo
authors.initials: SB
authors.lastname: Jiang
authors.forename: Shu-wen
authors.initials: SW
authors.lastname: Liu
mesh_terms: D016915:AIDS Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D000818:Animals; D002681:China; D015483:HIV Antibodies / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D016915:AIDS Vaccines; D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41; D022223:Vaccines, Subunit