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Reactivity of an HIV gag gene polypeptide expressed in E. coli with sera from AIDS patients and monoclonal antibodies to gag.
Marcus-Sekura CJ; Woerner AM; Klutch M; Quinnan GV Jr; Division of
June 30, 1988
Biochim Biophys Acta. 1988 Feb 28;949(2):213-23. Unique Identifier :

A segment of the gag gene of the human immunodeficiency virus (HIV) (HTLV-IIIB strain), the virus which causes acquired immunodeficiency syndrome (AIDS), has been cloned into the bacterial expression vector, pCQV2, and mapped to the right-hand portion of the gag gene containing the carboxyl-terminal portion of p24 and the amino-terminal portion of p15. Nucleic-acid sequencing of the insert-vector junctions further defined the 5'-terminal nucleotide of HIV sequence as nucleotide 997 and the 3'-terminal nucleotide as 1696. When used in an enzyme-linked immunosorbent assay (ELISA) with sera from HIV-infected patients, the cloned antigen reacted with a subset of sera which were positive on a standard ELISA using whole virus as antigen. Western-blot screening of these sera with whole virus indicated that all p24-positive sera were positive with the clone, suggesting that the carboxyl-terminal portion of p24 contains a highly antigenic epitope(s). A serum which was p24-negative p15-positive by Western blot analysis was also highly reactive, indicating that a p15 epitope is present in the cloned antigen. Epitope mapping with a series of monoclonal antibodies to gag resulted in positive ELISA with 2 of 3 anti-p24, 0 of 1 anti-p15, and 0 of 1 anti-p17 Western-blot-positive monoclonal antibodies, suggesting that one of the anti-p24 monoclonal antibodies reacts with epitopes amino-terminal to those coded from nucleotide 997, two anti-p24 monoclonals react with epitopes carboxyl-terminal to those coded from nucleotide 997, and the anti-p15 monoclonal reacts with epitopes carboxyl-terminal to those coded from nucleotide 1696.

Acquired Immunodeficiency Syndrome/*IMMUNOLOGY/MICROBIOLOGY Antibodies, Monoclonal/IMMUNOLOGY Antibodies, Viral/*IMMUNOLOGY Antigens, Viral/*GENETICS Cloning, Molecular Enzyme-Linked Immunosorbent Assay Epitopes Human HIV/*GENETICS/IMMUNOLOGY HIV Seropositivity/*IMMUNOLOGY Immunosorbent Techniques Retroviridae Proteins/GENETICS/*IMMUNOLOGY JOURNAL ARTICLE