Gateway Reporter Gene Fusions

Ben Holt bholt at email.unc.edu
Thu Oct 30 14:32:02 EST 2003


<x-flowed>Hello,
     I would like to get a consensus opinion on promoter fusions to
reporter genes, such as GUS and GFP, using GATEWAY modified binary
vectors. Are your labs making transcriptional or translational fusions?
The issues I see are 1) transcriptional fusions utilizing the repoter
gene ATG change the spatial relationship between your promoter and it's
endogenous ATG, and 2) translational fusions using the endogenous ATG
will add ~10 amino acids (from the recombination site) on the N-terminal
portion of the reporter gene and we don't know if this effects reporter
function. Anyone out there with experimental insights on this subject? I
would especially like to know whether anyone has done direct comparisons
between the methods, but any thoughts, ideas, etc. are greatly appreciated.
     I will be happy to compile the results for the entire group if I get
some responses.

Thanks a lot,
Ben

-- 
Ben Holt, PhD
University of North Carolina
Department of Biology
108 Coker Hall, CB#3280
Chapel Hill, NC 27599
(919)962-5838 - Lab#
(919)962-1625 - Fax
bholt at email.unc.edu


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