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Difference in Strains for Two Hybrid System

Megan Brown mbrown at fred
Tue Feb 14 20:07:03 EST 1995


I don't know about the difference between the two strains, but you could 
save yourself a lot of worrying by simply testing the 
interaction between the two proteins in vitro. In our 
lab, we do this routinely after detecting an 
interaction with the two-hybrid system. In addition, 
most journals will require an vitro demonstration of 
interaction before accepting your work for 
publication.

WANGHO at UCBEH.SAN.UC.EDU wrote: : Hi there!

: I sent the following mail last week, titled with "Difference between 2
: strains", but there is no comment or suggestion back.  If you read it before,
: please forget it.  I re-post it with a new name for people in Two hybrid system
: paying attention to it.   Thanks. 



: Re:  What is the difference between yeast strain ?

: Hi There!

: I am a newer in yeast but working on the yeast two hybrid system for almost
: three years.  A lot of exciting data gave me a big picture about the cross
: talking among the proteins I am interested in.  But one interaction bother me
: for a while.  I guess this might be a potential problem or could be a
: commo-problem in the future using yeast two hybrid system.

: 1. I detected the interaction between Protein A and Protein B in Yeast strain,
: PCY2.  The b-galactosidase activity induced by this interaction is beatiful.

: 2. Since some other strains have been developed (esp. using his and gal for
: double selection), I tried one.  All positive are positive,and all negative are
: negative.  Most interaction in my work showed positive.  But A-B interaction
: showed negative (both in his and b-galactosidase).

: 3. I made a conclusion that there is no direct A-B interaction but a complex
: containing A and B is formed, and the A-B complex is medicated by other
: factors.

: Above is the experiment and conclusion.  It appears to be perfect to tell this
: story in my next publication.  But, as I said, I am an user not a "yeastist". 
: So I basically want ask myself and "yeastists" three questions.

: 1. Is the difference between strains enough to giving these difference? I
: understand that the difference between two animal cells might be trillion.  Can
: some one tell me at what level the yeast strains show difference?

: 2. Am I so lucky to find this complex but an indirect interaction by using the 
: second strain, which I pick randomly "one day"?  

: 3. Obiviously, one like to know what is the third factor(s) mediating this
: interaction.  Any suggestion to provide that there is such factor(s) before I
: screening the yeast library (beacuse the screen is not the first choice, at
: least for me)?  Or is it easy to do substraction between this two strain?  A
: particular question is what is the regular size for a yeast cDNA library? 

: Any suggestion or comments are appreactiated.  If this is a true problem in the
: using of two hybrid system, I wish this should be paid attention to.  The
: false-positive might means more than what we thought.

: By the way, I can read "yeast at net.bio.net" through gopher.  You can send mail  
: to me or "yeast at net.bio.net".

: Thanks again!

: Hong F. Wang
: Department of Cell Biology, Neurobiology, and Anatomy
: University of CIncinnati College of Medicine
: 231 Bethesda Ave.
: Cincinnati, OH 45267-0521

: Phone: (513)558-6787
: Fax: (513)558-4454
: E-mail: wangho at ucbeh.san.uc.edu



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