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J Mol Biol. 2000 Feb 18;296(2):497-508.

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

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  • 1Istituto di Ricerche di Biologia Molecolare P. Angeletti, Via Pontina Km 30.600;, Roma, 00040 Pomezia, Italy.

Abstract

We describe the construction and characterization of two lambda surface displayed cDNA expression libraries derived from human brain and mouse embryo. cDNA inserts were obtained by tagged random-priming elongation of commercially available cDNA libraries and cloned into a novel lambda vector at the 3' end of the D capsid protein gene, which produced highly complex repertoires (1x10(8) and 2x10(7) phage). These libraries were affinity selected with a monoclonal antibody against the neural specific factor GAP-43 and with polyclonal antibodies that recognize the EMX1 and EMX2 homeoproteins. In both cases rapid identification of specific clones was achieved, which demonstrates the great potential of the lambda display system for generating affinity selectable cDNA libraries from complex genomes.

Copyright 2000 Academic Press.

PMID:
10669604
[PubMed - indexed for MEDLINE]
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