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Chromosome Two Polymorphisms

Martha K Jensen mkjensen at iastate.edu
Mon Nov 14 15:05:27 EST 1994


I'd like to thank everyone who wrote to tell me about their 
problems with the Wageningen tester line W6, and I hope that
this information can help other people avoid the frustration 
and delay that we all have experienced.

It seems that the gene as was isolated in Columbia ecotype
A. thaliana, and was crossed into a Landsberg line to create 
W6 (also known as CS6) containing cp2 ER as cer8.  The result of 
this is a Columbia island surrounding asymmetric, stretching
most of the way to ER.



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I have used an aux1-1, er line to do fine structure mapping around aux1. 
As far as I can tell this line is landsberg between marker 17288, 6191 and
4514/1789 (which are partially overlapping clones).  Further, the stock
center has an er py line which also gives landsberg recombinants in this
region.  I can send you information from my thesis, or information on
ordering a copy of my thesis from Indiana University.  Unfortunately aux1
was cloned by tagging in another laboratory prior to my reaching the locus
by walking, so my genetics data has yet to see the light of day.  I have
also constructed several strains for mapping in this region, including an
aux1, py, er triple mutant which I or Mark Estelle could send you.  as has
always been very difficult for me to score, aux1 is much easier, permittin
g
scoring as very young seedlings.  py is also very easy to score in my
hands.

Let me know if I can help,

Bryan

Bryan Pickett     Pickett at molbio.uoregon.edu   phone  503-346-4256
                                               fax    503-346-5891
Institute of Molecular Biology
University of Oregon
Eugene  OR  97403

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There is only one SSLP (microsatellite Marker) in that region with
the name nga168. It will give with Col a 151 bp fragment and with Ler
a 135 pb fragment.
The protocol is published Bell and Ecker 1994: Genomics 18, 137-144
Perhaps you can find a other polymorphism. But propably you should
redo the cross and use a normal Ler parent.
You could also try the AFLP method to find a new polymorphic marker
between you mutant and the wild type. Hopefully I will get in the
near future a reliable protocol for AFLP's.
Good luck
Marie-Theres Hauser

Center of Applied Genetics
Universitaet fuer Bodenkultur
Gregor Mendel Str. 33
A-1180 Wien
Austria

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I & Bryan Pickett mapped aux1 (also in that region) but had the same problem
you did with that mapping line. I think that the as marker originally came from
Columbia. I recommend using er, also aux1-1 is in a Ler background so you can
use aux1-1 er double to cross to your gene & if its between the two genes
you can select tons of recombinants by identifying single mutants in the F2
(this is the scheme we used).
--Lawrence Hobbie (estelle lab)

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28 October 1994

Dear Martha

We also had difficulties with RFLP mapping in the region between as and er.  Li
ke you, we 
assumed that the asymmetric leaves mutant in the Waginengen tester line was iso
lated in 
Landsberg erecta background.  When it came to the point, all RFLP markers in th
e vicinity of 
asymmetric leaves were clearly Columbia.  So we looked up the origin of asymmet
ric leaves 
in the Caltech list of mutants (the green book of Meyerowitz and Pruitt), and, 
sure enough, it 
was isolated by Redei and Hirono (1964) Arabidopsis Information Service Vol 1, 
pages 9-10.  
Redei generated the Columbia wild type, and used it extensively for his mutant 
hunts.

So, I think that's your likely answer.

Cheers,

David Smyth
==========================================================
David Smyth
Department of Genetics and Developmental Biology
Monash University
Clayton, Vic. 3168
Australia                     

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