I have a question concerning use of the two-hybrid system to look for
proteins which interact in a calcium DEPENDENT manner.
We would like to identify proteins which interact with a calcium-binding
protein, but the question has been raised as to the nuclear content of
calcium in yeast cells and if there is any manner in which calcium levels
can be increase intracellularly. We have considered growth of the yeast on
calcium chloride containing plates, but we are not sure whether the yeast
will sequester the additional calcium or if it will alter the steady state
levels of calcium.
Has anyone considered this problem further who might have a solution?
Please post responses to me, to alleviate clutter on the net.
kaj at bioc.rice.edu