A versatile system for the production of recombinant chimeric peptides.
Production of chimeric and multimeric peptides is of interest for the analysis of topographic relationships between T and B cell stimulatory epitopes. Recombinant DNA technology has certain advantages over conventional chemical peptide synthesis for the production of peptide constructs of large size (more than 40 amino acid residues). We describe a methodology which is versatile and independent of the expression vector used because it only relies on the incorporation of appropriate restriction enzyme sites in oligonucleotides. The method was verified using two 20mer sequences from the 38 kDa antigen of M. tuberculosis. Peptide 201-220, containing an antibody binding linear epitope, has been made immunogenic in vivo when combined with T cell stimulatory peptide 350-369 in a chimeric peptide. The results demonstrate that a distinct orientation of the constituent peptides was essential for achieving optimal immunogenicity.
De Smet
K A
KA
Tuberculosis and Related Infections Unit, MRC Clinical Sciences Centre, Hammersmith Hospital, London, UK.
Vordermeier
H M
HM
Ivanyi
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000942:Antigens, Bacterial / Q000737:chemistry*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D000939:Epitopes; D005260:Female; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000276:immunology; D010455:Peptides / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D000942:Antigens, Bacterial; D000939:Epitopes; D010455:Peptides; D011993:Recombinant Fusion Proteins