Fig. 6. Minimal eIF2Bε^518–712 fragment provides nucleotide exchange activity in vivo. (A) growth of strains GP4113 (left) and GP4115 (right) on YPD agar. Photographed after 5 days at 30°C (B). Western blot analysis of GP4113 (lane 1) and three 5-FOA-resistant colonies (lanes 2–4, where GP4115 is lane 2). Fifteen micrograms of total soluble cell protein was loaded in each lane. Blot was probed with the specific antisera indicated to the right of each panel. (C) Growth of indicated transformants of strain GP4115 on SCGR minus uracil, leucine, isoleucine and valine (SCGR-ULIV) media. ‘Vector’ indicates plasmid pRS316 (Sikorski and Hieter, 1989) and ‘GCD6’ the low copy GCD6 URA3 plasmid pJB85 (Bushman et al., 1993); other plasmids are indicated in Figure 1. Times listed are doubling times ± SD (σn) for each strain in liquid SCGR-ULIV during exponential growth phase. (D) Growth of indicated transformants of strain GP4181 on SCGR-uracil media. ‘GCD2’ indicates the low copy GCD2 URA3 plasmid pCP62 (Pavitt et al., 1997).