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An alkali-sensitive mutant, 18224, of the alkaliphilic Bacillus sp. strain C-125 was characterized. The nucleotide sequence of the PvuI-NlaIV DNA fragment that recovers the alkaliphily of 18224 has been cloned from the mutant and sequenced. Comparison of the nucleotide sequences of the corresponding regions found a G to A substitution in the mutant. The mutation resulted in an amino acid substitution from 82Gly to Glu of the putative ORF3 product, which consisted a gene cluster of at least four tandemly located open reading frames. The ORF3 product was deduced to be an 112 amino acid polypeptide with hydrophobic properties, which was expressed using an in vitro translation system.
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