The Expression in Escherichia coli of Sequences Coding for the p18 Protein of Human Immunodeficiency Virus and the Use of the Recombinant Protein in Characterizing a Panel of Monoclonal Antibodies against the Viral p18 Protein

Abstract

SUMMARYThe sequences coding for the pl8 protein of CBL-1, a British human immunodeficiency virus (HIV) type 1 isolate, were expressed in Escherichia coli as fl-galactosidase fusion proteins. The recombinant proteins were used to screen a panel of five monoclonal antibodies (MAbs) raised against pl 8 expressed in CBL-l-infected cells. The regions containing the epitopes for four of the MAbs were located using carboxy deletion mutants and synthetic peptides. The epitope of one of the MAbs (1D9) was reconstructed as part of an unfused, E. coil-expressed p 18 protein using the polymerase chain reaction technique. Four different HIV strains and one lymphadenopathy virus type 2 strain were analysed by fluorescence-activated cell sorting of live infected cells using the plS-reactive MAbs.

The Cloning and Expression in Escherichia coli of Sequences Coding for p24, the Core Protein of Human Immunodeficiency Virus, and the Use of the Recombinant Protein in Characterizing a Panel of Monoclonal Antibodies against the Viral p24 Protein

“…In the accompanying paper (Spence et al, 1989) we describe the expression of the pl8 protein and the analysis of different HIV isolates using the anti-pl8 MAbs.…”

The role of proteolytic processing in the morphogenesis of virus particles

“…Morphogenesis resembles that of bacteriophages in that assembly is dependent on virusencoded scaffolding proteins that are eliminated from the mature capsid 13'24, and in that maturation involves extensive assembly-dependent proteolysis of capsid proteins. At least six virion polypeptide precursors are cleaved to mature components by a 23 kDa virus-encoded proteinase (references in Webster et al 116) after assembly of immature virions and subsequent encapsidation of DNA. Processing is presumably regulated to prevent premature cleavage of precursors before assembly, but the mechanism by which this is achieved has not been elucidated.…”

Expression of feline immunodeficiency virusgag gene inEscherichia coli

“…Spence et al characterized a panel of monoclonal antibodies against viral core protein 384 T. Furuya et al [14,15], and Ghrayeb et al detected anti-gag antibodies in sera 1-2], using the gag proteins expressed in Escherichia coli. Jacobs et al showed that the gag precursor protein synthesized in yeast is myristoylated and targeted to the plasma membrane [4].…”