[Yeast] His3 background test

[Valiveti, Aswani]

Hi,

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?

Will greatly appreciate any helpful advice.

Thanks for your attention.

regards,
Aswani.
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