PET56 clone

Colin MacD macdiarm at sbsu1.auckland.ac.nz
Wed Aug 2 18:57:12 EST 1995


Hi out there
I have run into a problem which I would appeciate some help with.  I am using
a PCR gene disruption method (oligos which amplify the his3 gene to give a product with 50 bp homology to the target gene at each end).  This method requires the use of a his3 deletion strain (to reduce integration at the his3 locus).  I am using his3D200 allele, but found recently that this is associated with a petite phenotype (since the his3 deletion removes some of the promotor of the adjacent pet56 gene) - so if I use a diploid his3D200 strain I cant sporulate it!  Has anyone else come across this problem? 
Anyway, I am going to try  the disruption in the diploid, then try putting the pet56 gene in on a plasmid.  Does anyone have a clone of the pet56 gene?  Would much appeciate the gift if it is available.
Thanks in advance,

Colin MacDiarmid
macdiarm at sbsu1.auckland.ac.nz



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