IUBio

Saccharomyces expression!!!!

Pathology small_group.pathology at MUWAYE.UNIMELB.EDU.AU
Thu Jun 27 01:25:38 EST 1996


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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