Capturing reaction paths and intermediates in Cre-loxP recombination using single-molecule fluorescence.

ABSTRACT: Site-specific recombination plays key roles in microbe biology and is exploited extensively to manipulate the genomes of higher organisms. Cre is a well studied site-specific recombinase, responsible for establishment and maintenance of the P1 bacteriophage genome in bacteria. During recombination, Cre forms a synaptic complex between two 34-bp DNA sequences called loxP after which a pair of strand exchanges forms a Holliday junction (HJ) intermediate; HJ isomerization then allows a second pair of strand exchanges and thus formation of the final recombinant product. Despite extensive work on the Cre-loxP system, many of its mechanisms have remained unclear, mainly due to the transient nature of complexes formed and the ensemble averaging inherent to most biochemical work. Here, we address

SUBMITTER: Pinkney JN 

PROVIDER: S-EPMC3529024 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature