Cloning Aspergillus fumigatus allergens by the pJuFo filamentous phage display system.

Abstract

A cDNA library from Aspergillus fumigatus has been displayed on the surface of filamentous phage M13 and screened for gene products binding to human serum IgE. The physical linkage of cDNA gene products to the genetic information required for their production, achieved by exploiting the high-affinity interaction of the Jun and Fos leucine zippers, allows rapid and easier screening of large libraries in semifluid systems. The pJuFo cloning vector is designed to display proteins on the surface of phage and allows selective isolation of genes by gene product-ligand interaction. Thus the system is applicable to clone cDNA that encodes proteins for which a ligand is available. Herein we show that phage expressing IgE binding proteins from A. fumigatus can be enriched and separated from nonspecific phage by using serum IgE from A. fumigatus-allergic individuals coated to plastic dishes. Subsequently, the proteins can be produced in high amounts in Escherichia coli and purified for usage in allergy testing.