Cloning and Sequencing of the gerD Gene of Bacillus subtilis

School of Biological Sciences, University of Birmingham, Birmingham B15 2TT, UK ABSTRACT Summary: A Tn 917 insertion in the same region of the chromosome as gerD gave rise to a mutant ( ger-97 ) with a germination phenotype similar to that of two gerD mutants which germinate abnormally in a range of germinants. The insertion and two gerD mutations were cotransformed with ribosomal protein genes rpoB, rpsE and rpsl. DNA cloned from one side of the insertion carried the 16S end of the ribosomal RNA operon rrnl. These data were consistent with the order rpoB-rpsE-rpsI-gerD/ger-97 :: Tn917-rrnI. Insertion into the wild-type chromosome of a plasmid carrying DNA adjacent to the insertion permitted the recovery of a 1·8 kb fragment of DNA which complemented ger-97: :Tn9/7and the gerD mutations. The DNA nucleotide sequence of the region of this fragment at which Tn917 had inserted revealed a 555 bp open reading frame, preceded by a ribosome-binding site and potential E and A promoter regions and encoding a predicted polypeptide of 21117 Da. This polypeptide was largely hydrophilic but contained a hydrophobic region at the N-terminus resembling a signal peptide. Present address: Imperial Cancer Research Fund, Lincolns Inn Fields, London WC2A 3PX, UK. Present address: Department of Anatomy, University of Birmingham, Birmingham B15 2TJ, UK.