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Requesting genomic clones from all arabinetters

Tue Aug 30 10:59:45 EST 1994

To all members of the Arabidopsis community,

I am a postdoc in Dr. Craig Pikaard's lab at Washington University, and I 
am appealing to all Arabinetters to help in my search hormone responsive 
genes and ribosomal protein genes.  My research is on the regulation of 
rRNA gene transcription, and associated processes.  I am currently 
investigating the response of the rRNA gene and spacer promoters to 
hormones.  So far my work has proved interesting, but in order to 
quantitate the effects of the various hormone treatments I need to 
normalize my results to an internally controlled standard, non-hormone 
responsive gene.  Or at least to genes that are known not to respond 
directly or indirectly to certain classes of hormones.  I also need 
genes that are known to respond to the various hormone classes such that I 
have a positive control for the hormone treatment.  It wouldn't do to say 
that rRNA genes don't respond to a certain hormone if I couldn't also show 
that genes known to respond to that hormone show an induction.  I also 
would like to obtain ribosomal protein genes so that I could use them as 
internal controls.  
Since my experiments involve S1 protection analysis and Nuclear run-on 
transcription, I need genomic clones so that the entire gene from the start 
site of the promoter through at least the first 500 or so base pairs 
including intron sequences.  I would also like pertinent restriction sites 
or sequence information.  I would greatly appreciate any help members of 
the Arabidopsis community could provide.  
Additionaly, I have had a string of sucessful nuclear run-on transcription 
reactions.  The only real problem that I am encountering is recovery of all 
the nuclei.  I get back only about 10 to 20% of the total nuclei liberated 
by cell disruption.  However the nuclei are extremely active.  I am 
currently working on ways to increase the overall yield of nuclei, and when 
I do so I will make the entire procedure (from isolation of nuclei to 
hybidization of the labeled RNA) available on the network.

Thank you in advance,
					Reggie Gaudino
Reggie Gaudino			Internet:     gaudino at wustlb.wustl.edu
				BITnet:       GAUDINO at WUSTLB
Washington University		Biology Department  Box 1137
St. Louis, Missouri  63130	(314)935-8529

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