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Saccharomyces expression!!!!

Hoffmacs Hoffmacs at HERMES.BC.EDU
Thu Jun 27 04:58:19 EST 1996


Dear Gian et al.,
I think one is a contaminant.  I suggest two things.
1.  Look at them under a microscope.  Also look at the parent strain.
2.  Check for growth on other defined media to score the other auxotrophies
in your host strain.  
My money is on the smaller colonies.  Good luck.
Charlie Hoffman
"I'd rather be fission."
________________________________________________________

To whoever can help me,

I have recently obtained the yeast expression vector YEpFLAG-1 (from Kodak)
and cloned my gene of 
interest into it and then electroporated this into the host strain BJ 3505. 
When I checked my colonies 
a few days later (on a SCM -trp plate), I noticed 2 types of colonies on the
plate;
(1) rather large, cream colored, broad colonies and;
(2) smaller, white-colored, well defined colonies.

When I grew these colonies in some SCM -trp media, the cream colonies grew
quite well, while the white 
colonies took an extra day or two to grow up (yeast containing no vector
didn't grow at all-neg. 
control).  When grown in a complete media containg peptone, glucose,
glycerol, etc..., the cream 
colonies only grew to a low cell density, while the white colonies grew to a
higher density.

If anyone out there has used the YEpFLAG1 vector or has any ideas what is
going wrong(?!?!) with this, 
could you please contact me via e-mail on;
small_group.pathology at muwaye.unimelb.edu.au

Thanking you for your time and assistence,

Gian Sberna
Dept. of Pathology,
University of Melbourne,
Parkville, Victoria
Australia, 3052.





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