Document 1007
 DOCN  M94A1007
 TI    Molecular characterization of a HIV-1 strain (HIV-1IbNg) from Nigeria.
 DT    9412
 AU    Howard TM; Rasheed S; Laboratory of Viral Oncology and AIDS Research,
       University of; Southern California 90032-3626.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(2):23 (abstract no. 384A). Unique
       Identifier : AIDSLINE ICA10/94371568
 AB    OBJECTIVE: To characterize the overall genomic organization of a new
       strain of HIV-1 (HIV-1IbNg) from Nigeria. METHODS: HIV-1IbNg was
       isolated from the peripheral blood mononuclear cells (PBMC) of a
       Nigerian individual using coculture techniques and
       phytohemagglutinin-stimulated PBMC from a HIV seronegative donor. Two
       different techniques were then employed to molecularly clone this virus
       from infected PBMCs. Firstly, the RT-PCR technique (reverse
       transcriptase, polymerase chain reaction) was used to amplify the
       HIV-1IbNg genome using total, cytoplasmic RNA and resultant amplicons
       were cloned. Secondly, a phage library was constructed using
       unintegrated, circular viral DNA extracted by the Hirt procedure from
       infected PBMCs. Recombinant phage containing the HIV-1IbNg genome were
       identified using a probe generated from one of the recombinant plasmids
       containing the pol/env coding region of the genome. All DNA sequencing
       was done by the dideoxy-chain termination method using double-stranded
       DNA templates. RESULTS: A comparison of the nucleotide sequences of the
       gp120 coding regions of HIV-1IbNg and other HIV-1 strains revealed that
       this virus is most closely related to strains of HIV-1 belonging to the
       'A' subtype. Significant differences in all five hypervariable regions,
       especially the V4 region, was noted when the deduced amino acid sequence
       of the gp120 proteins of HIV-1IbNg and other viral strains grouped in
       clade 'A' were compared. The number and positions of the cysteine
       residues along with the number and positions of potential
       N-glycosylation sites, however, were found to be highly conserved
       amongst these strains. DISCUSSION AND CONCLUSIONS: This is the first
       known isolate of HIV-1 of Nigerian origin which has been molecularly
       cloned and characterized. Based on information obtained from the env
       region of the viral genome, HIV-1IbNg clusters most closely with strains
       of HIV-1 of the 'A' subtype which have been isolated from inhabitants of
       central and western Africa. Characterization of HIV-1IbNg is important
       for the development of vaccines and diagnostics which may be used in
       regions of the world where HIV-1 strains of the 'A' subtype are endemic.
 DE    Cloning, Molecular  Comparative Study  Genes, env  *Genome, Viral  Human
       HIV Envelope Protein gp120/GENETICS  HIV Infections/MICROBIOLOGY
       HIV-1/CLASSIFICATION/*GENETICS/ISOLATION & PURIF  Leukocytes,
       Mononuclear/MICROBIOLOGY  Nigeria  Polymerase Chain Reaction
       Viremia/MICROBIOLOGY  MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).