Fig. 4. Cln2 performs important roles in both the nucleus and the cytoplasm. (a) An empty control vector (pRS315), a low-copy plasmid containing CLN2 with no tag (CLN2/pCB1314) and plasmids containing CLN2 with C-terminal forced localization cassettes under control of the CLN2 promoter (pNE10X series) were transformed into strain N-138 (cln1 cln2 cln3 GAL-CLN1). Cells were serially diluted (see Materials and Methods) onto selective plates containing 2% galactose (GAL) or glucose (GLC), and grown for 2 days at 30°C. Within a single dilution series, the number of cells in each spot differs by tenfold from its neighbor (b) An empty control vector (pRS315), a low copy plasmid containing CLN2 with no tag (CLN2/ pNE113) and plasmids containing CLN2 with C-terminal forced localization cassettes under control of the methionine-repressible MET3 promoter (pM10X series) were transformed into strain YHW23 (swi4 swi6 GAL-SWI4). Strains were serially diluted as in (a) on plates containing 2 mM methionine or lacking methionine (–MET) and containing 2% galactose or glucose. (c) The same plasmids were used as in (b), but were transformed into strain N-162 (swi4 swi6 GAL-SWI4 sic1), and cells were grown for 5 days at 30°C.
