A NOVEL PHAGE-LAMBDA REPLACEMENT CRE-LOX VECTOR THAT HAS AUTOMATIC SUBCLONING CAPABILITIES

We have developed a novel phage lambda replacement cloning vector, lambdapAn. LambdapAn allows one to automatically subclone the insert as a plasmid using the Cre-loxP site-specific recombination system. This eliminates the need to subclone insert fragments and permits the rapid structural analysis of insert DNA. LambdapAn is similar to other phage lambda replacement vectors taking inserts ranging in size from 5 to 19 kb. We have placed the pyrG gene of Aspergillus nidulans on the vector as a nutritional selective marker for transformation. We have developed this vector as part of an overall plan to facilitate the cloning of dominant extragenic suppressor mutations from A. nidulans, but also know that it is a generally useful vector for the purposes of isolating genomic clones without the need to subclone from the phage lambda vector.