Recombineering-mediated tagging of Drosophila genomic constructs for in vivo localization and acute protein inactivation.

ABSTRACT: Studying gene function in the post-genome era requires methods to localize and inactivate proteins in a standardized fashion in model organisms. While genome-wide gene disruption and over-expression efforts are well on their way to vastly expand the repertoire of Drosophila tools, a complementary method to efficiently and quickly tag proteins expressed under endogenous control does not exist for fruit flies. Here, we describe the development of an efficient procedure to generate protein fusions at either terminus in an endogenous genomic context using recombineering. We demonstrate that the fluorescent protein tagged constructs, expressed under the proper control of regulatory elements, can rescue the respective mutations and enable the detection of proteins in vivo. Furthermore, we also a

SUBMITTER: Venken KJ 

PROVIDER: S-EPMC2566861 | biostudies-literature | 2008 Oct

REPOSITORIES: biostudies-literature