A Mutational Analysis of the Bacteriophage P1 Recombinase Cre

Abstract

Bacteriophage P1 encodes a 38,000 dalton site-specific recombinase, Cre, that is responsible for reciprocal recombination between sites on the P1 DNA called loxP. Using in vitro mutagenesis 67 cre mutants representing a total of 37 unique changes have been characterized. The mutations result in a wide variety of phenotypes as judged by the varying ability of each mutant Cre protein to excise a lacZ gene located between two loxP sites in vivo. Although the mutations are found throughout the entire cre gene, almost half are located near the carboxyl terminus of the protein suggesting a region critical for recombinase function. DNA binding assays using partially purified mutant proteins indicate that mutations in two widely separated regions of the protein each result in loss of heparin resistant complexes between Cre and a loxP site. These results suggest that Cre may contain two separate domains both of which are involved in binding to loxP.