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Two-hybrid interaction between Arc35p and Cmd1p. ARC35 and arc35-1 were cloned in frame to the lexA DNA binding domain of pEG202. CMD1 and the collection of ts cmd1 alleles were cloned in frame behind the B42 transactivation domain of pJG4-5. β-Galactosidase activity indicative of interaction was monitored on X-Gal plates at 37°C supplemented with no calcium or 5 mM calcium.
