Open AccessA peptide library expressed in yeast reveals new major epitopes from human immunodeficiency virus type 1
Author(s) -
Madaule Pascal,
Gairin Jean Edouard,
Bénichou Serge,
Rossier Jean
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04203.x
Subject(s) - epitope , biology , monoclonal antibody , virology , microbiology and biotechnology , gp41 , recombinant dna , peptide library , plasmid , restriction enzyme , cdna library , complementary dna , antibody , peptide sequence , dna , gene , genetics
Summary In order to characterize novel human immunodeficiency virus type 1 (HIV‐1) continuous epitopes, we designed a simple method, based on recombinant DNA, providing a complete set of peptides derived from HIV‐1. A library (4 × 10 4 clones) was first constructed in a new expression/secretion vector, using as inserts small fragments of HIV‐1 DNA (50–150 bp) generated by random DNAse I cleavage. This peptide library, expressed in the yeast Saccharomyces cerevisiae , was screened with sera of HIV‐1 infected individuals and human and murine anti‐HIV‐1 monoclonal antibodies. Plasmids from immunoreactive colonies were recovered and the sequences of the HIV‐1 derived inserts were determined. By using human sera, we have detected classical HIV‐1 epitopes and identified two novel major epitopes, which may be used to improve diagnostic tests, localized in the p24 core protein and in the endonuclease. In addition, four minor epitopes were also defined by screening the library with monoclonal antibodies: in the protease, in the p17 core protein, in gp120 and near the C‐terminal of gp41. This method is general and can be used for any protein from which a cloned cDNA is available.