In article <7oq6gn$7t1 at net.bio.net>,
news at newsreader.wustl.edu, Alex Brands <abbrands at artsci.wustl.edu> wrote:
>> I would like to generate a null allele of my new favorite gene in yeast.
> The gene I'm interested in is very close to the neighboring gene, and they
> are oriented in opposite directions, such that there is only 234 bases
> between the start codons. I'm concerned that the promoter of the
> neighboring gene extends into the coding sequence of the gene I want to
> knock out. If this is true, if I knock out the whole coding sequence, I
> will also disturb normal expression of the neighboring gene.
>> With the tightly packed yeast genome, this must be a relatively common
> problem, how should I deal with it?
>> Thanks for any input,
>> Alex Brands
Why not knock out both ORFs and compare their phenotypes? Is the second
gene essential? Also, you could make a smaller deletion of your gene of
interest, so as to start comfortably downstream of the ATG, and then check
to see if you've generated a null via Northern and/or Western.
Ed Davis, Ph.D.
ABL-Basic Research Program
Sent via Deja.com http://www.deja.com/
Share what you know. Learn what you don't.
YEAST bionet newsgroup see: http://www.bio.net/hypermail/YEAST/
YEAST e-mail: messages sent to yeast at net.bio.net
subscribe: e-mail biosci-server at net.bio.net with: subscribe yeast
unsubscribe: e-mail biosci-server at net.bio.net with: unsubscribe yeast
YEAST on the WWW: http://genome-www.stanford.edu/Saccharomyces/VL-yeast.html
problems with the YEAST newsgroup? E-mail the moderator: francis at cmmt.ubc.ca