Construction of strains expressing the GFP-CHAP1 fusion protein. A. Construct map. GFP, GFP gene; Ptrp, constitutive promoter; HPH, hygromycin phosphotransferase gene; chap-m, chap-s, and gfp-f, primers used for transformant verification. Upper drawing: transformant; lower drawing: wild type. B. Confirmation of CHAP1-GFP expression by reverse transcription-PCR. The following reactions were performed with RNA of the wild type and transformants gfp-chap1 and gfp-chap2. Lanes 1, gfp-f and chap-s; lanes 2, chap-m and chap-s. Control reactions without reverse transcriptase (−RT) were performed with primers gfp-f and chap-s for transformants and chap-m and chap-s for the wild type. The predicted product size, 1.8 kb, is indicated at the left of the gel; note that this is smaller than the size indicated in A, 1,976 bp, due to splicing of introns.