RT Journal
A1 Hope, Ian A.
A1 Stevens, Jonathan
A1 Garner, Anna
A1 Hayes, Josie
A1 Cheo, David L.
A1 Brasch, Michael A.
A1 Vidal, Marc
T1 Feasibility of Genome-Scale Construction of Promoter::Reporter Gene Fusions for Expression in Caenorhabditis elegans Using a MultiSite Gateway Recombination System
JF Genome Research 
JO Genome Research 
YR 2004 
FD October 15 
VO 14 
IS 10b 
SP 2070 
OP 2075 
DO 10.1101/gr.2463804 
UL http://genome.cshlp.org/content/14/10b/2070.abstract 
AB The understanding of gene function increasingly requires the characterization of DNA segments containing promoters and their associated regulatory sequences. We describe a novel approach for linking multiple DNA segments, here applied to the generation of promoter::reporter fusions. Promoters from Caenorhabditis elegans genes were cloned using the MultiSite Gateway cloning technology. The capacity for using this system for efficient construction of chimeric genes was explored by constructing promoter::reporter gene fusions with a gfp reporter. The promoters were found to provide appropriate expression of GFP upon introduction into C. elegans, demonstrating that the short Gateway recombination site between the promoter and the reporter did not interfere with transcription or translation. The recombinational cloning involved in the Gateway system, which permits the highly efficient and precise transfer of DNA segments between plasmid vectors, makes this technology ideal for genomics research programs.