Bacteriophage lambda display of complex cDNA libraries: A new approach to functional genomics

We describe the construction and characterization of two lambda surface dis played cDNA expression libraries derived from human brain and mouse embryo. cDNA inserts were obtained by tagged random-priming elongation of commerci ally available cDNA libraries and cloned into a novel lambda vector at the 3' end of the D capsid protein gene, which produced highly complex repertoi res (1 x 10(8) and 2 x 10(7) phage). These libraries were affinity selected with a monoclonal antibody against the neural specific factor GAP-43 and w ith polyclonal antibodies that recognize the EMX1 and EMX2 homeoproteins. I n both cases rapid identification of specific clones was achieved, which de monstrates the great potential of the lambda display system for generating affinity selectable cDNA libraries from complex genomes. (C) 2000 Academic Press.