Immunoblot analysis of proteins encoded by the GALLS gene. Blots were probed with GALLS-specific antibodies. FL, GALLS-FL; His-FL, His6-tagged GALLS-FL; CT, GALLS-CT. Numbered bars indicate the size (in thousands) of molecular weight standards. (A) Soluble proteins from A. tumefaciens containing vector plasmid (left lane) or the wild-type GALLS gene (right lane). (B) Soluble proteins were extracted from A. tumefaciens cells harboring the His6-GALLS-FL gene and fractionated on Ni-nitrilotriacetic acid resin; bound proteins were eluted with imidazole. Proteins from the flowthrough (flow), wash, and first and second eluates are shown. (C) Soluble proteins from A. tumefaciens containing the vector plasmid (lane 1), wild-type GALLS gene (lane 2), the +1 frameshift mutation in codon 803 of the GALLS gene (lane 3, +1 FS), or the GALLS gene with the Met808Ile Met815Val mutations (lane 4, MIMV). (D) GALLS proteins expressed in A. tumefaciens cells containing the GALLS gene fused to the Trc promoter and 5′ UTR from pTrc99 in the absence (−) or presence (+) of IPTG.