Cloning, sequencing, and expression of the rpoD gene encoding the primary sigma factor of Xanthomonas campestris

A DNA fragment encoding the primary sigma factor from Xanthomonas campestris pv. campestris was cloned and sequenced. The gene (rpoD) encodes a polypeptide of 622 amino acids with a calculated MW of 70,700. The deduced amino acid sequence exhibits extensive sequence homology to the conserved regions of the primary sigma factors from bacteria. The gene product expressed in Escherichia coli, detected by Western blot analysis, had a MW similar to that estimated for the purified protein in SDS-PAGE. The NH2-terminal amino acid sequence determined chemically matched with that deduced from the nucleotide sequence of the rpoD gene. The calculated pI value (9.31) for the X. campestris primary sigma factor is much higher than the values observed for the analogous proteins from other bacteria.