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degradation of lexA fusion protein?

Françoise Martz Francoise.Martz at plantphys.umu.se
Thu Apr 8 15:15:08 EST 1999


I am doing control for the two-hybrid system to detect the lexA fusion
protein in yeast extracts by western blotting. I used too bait
constructs: a complete coding region and a 64aa-deleted one at the
N-term end. With a specific antibody, I detected a band of ca. 37 kDa
instead of the expected 78 kDa in the strain containing the complete
construct, and nothing in the strain containing the uncomplete
I used 2 different extraction protocols, both with PMSF and one with
leupeptin and  benzamidin as well.
I am not familar with the yeast world. Does someone already encounter
the same problem? What other control should/can I do? Can I perform the
two-hybrid screen? 
And where can I test my protein sequence for protease cleavage sites?

Thanks a lot for your help....


Francoise.Martz at plantphys.umu.se

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