FIG. 5.

Detection of YaaH-His in sporulating cells. (A) pYAAH8 has replication origins available in both E. coli (ori-E) and B. subtilis (ori-B) cells, and it confers tetracycline resistance on these organisms. The yaaH gene in pYAAH8 is regulated by a promoter located upstream of the gene and fused to a sequence encoding six consecutive histidine residues (His tag). (B) B. subtilis 168 was transformed with control vector pTUBE1200H6 or pYAAH8 as described in Materials and Methods. Proteins prepared from the transformants were resolved by SDS-PAGE (15% acrylamide gel) and visualized by immunoblotting with antisera against the His tag and SigK. Arrowheads indicate the positions of the YaaH-His protein and SigK. T, harvesting times for cells (in hours).