A toolkit and robust pipeline for the generation of fosmid-based reporter genes in C. elegans.

ABSTRACT: Engineering fluorescent proteins into large genomic clones, contained within BACs or fosmid vectors, is a tool to visualize and study spatiotemporal gene expression patterns in transgenic animals. Because these reporters cover large genomic regions, they most likely capture all cis-regulatory information and can therefore be expected to recapitulate all aspects of endogenous gene expression. Inserting tags at the target gene locus contained within genomic clones by homologous recombination ("recombineering") represents the most straightforward method to generate these reporters. In this methodology paper, we describe a simple and robust pipeline for recombineering of fosmids, which we apply to generate reporter constructs in the nematode C. elegans, whose genome is almost entirely covered

SUBMITTER: Tursun B 

PROVIDER: S-EPMC2649505 | biostudies-literature | 2009

REPOSITORIES: biostudies-literature