Yersinia enterocolitica cells, when cultured at 30 degrees C or below, are flagellated and motile. Cells cultured at 37 degrees C or above lack flagella and are non-motile. To identify flagellin genes that are a target of this temperature-dependent regulation, a library of Y. enterocolitica genomic inserts in a phage lambda vector was probed with the Salmonella typhimurium fliC (flagellin) gene. A DNA fragment subcloned from a recombinant phage which hybridizes with the probe complements a non-motile S. typhimurium fliC-fljB- (flagellin-minus) mutant. DNA sequence analysis shows that Y. enterocolitica contains three tandem flagellin genes, designated fleA, fleB and fleC. All three genes are co-ordinately transcribed at low, but not high, temperature from fliA-dependent (sigma F) promoters. Flagellin transcription arrests rapidly after upshift to 37 degrees C (host temperature). In contrast, flagellin transcription resumes only after several generations when cells cultured at 37 degrees C are downshifted to 28 degrees C.