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<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2>Hi,</FONT></SPAN></DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial size=2>I am performing a
one hybrid experiment using pHisi-1. pLaczi reporter vectors and pJG4-5-cDNA
library in EGY48 strain. While testing for background His3 reporter expression I
find that the yeast grow even at 125mM 3-AT. The strains with plasmid alone also
grow at 60mM 3-AT, further concentrations not tested yet. How high concentration
of 3-AT can I use to completely eradicate the growth due to leaky expression.
What are the effects of 3-AT on yeast growth, aside from Histidine biosynthetic
pathway? Will changing the yeast host strain help in reducing the
background?</FONT></SPAN></DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial size=2>Will greatly
appreciate any helpful advice.</FONT></SPAN></DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial size=2>Thanks for your
attention.</FONT></SPAN></DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2>regards,</FONT></SPAN></DIV>
<DIV><SPAN class=672401019-28012008><FONT face=Arial
size=2>Aswani.</FONT></SPAN></DIV></BODY></HTML>
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