A gene, , encoding a spore coat polypeptide of , was isolated from an fragment (54 kb) of the chromosome by using synthetic oligonucleotide probes corresponding to the NH-terminal amino acid sequence of Cot40-2 previously purified from the spore coat of The nucleotide sequence (2603 bp) was determined and sequence analysis suggested the presence of two contiguous ORFs, ORF X and , followed by the 5′-region of an additional ORF, ORF Y, downstream of The gene is 1053 nucleotides long and encodes a polypeptide of 351 amino acids with a predicted molecular mass of 41083 Da. The predicted amino acid sequence was in complete agreement with the NH-terminal amino acid sequence of Cot40-2. The gene is 1131 nucleotides long and encodes a polypeptide of 377 amino acids with a predicted molecular mass of 42911 Da. The gene product of was confirmed to be identical to Cot40-2 by SDS-PAGE and immunoblotting from transformed with a plasmid containing the region. Northern hybridization analysis indicated that a transcript of and appeared at about 5 h after the onset of sporulation. The transcriptional start point determined by primer extension analysis suggested that −10 and −35 regions are present upstream of and are very similar to the consensus sequence for the -dependent promoter. Terminator-like sequences were not found in the DNA fragment (2603 bp) sequenced in this paper, which suggested that the locus may be part of a multicistronic operon. The gene is located between and at about 270-275° on the genetic map. Insertional mutagenesis of the gene by introducing an integrative plasmid resulted in no alteration of growth or sporulation, and had no effect on germination or resistance to chloroform.


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