1887

Abstract

On the basis of the genome, a novel type of operon was deduced. The gene expression and biochemical properties of this operon were further characterized. RT-PCR analysis of the intergenic regions suggested that the transcription of the operon was continuous. With gene cloning and enzyme activity assays, TTE1929, TTE1928 and TTE1927 were identified to be GalT, GalK and GalE, respectively. Results elicited from polarimetry assays revealed that TTE1925, a hypothetical protein, was a novel mutarotase, termed MR-Tt. TTE1926 was identified as a regulator that could bind to two operators in the operon promoter. The transcriptional start sites were mapped, and this suggested that there are two promoters in this operon. Expression of the genes was significantly induced by galactose, whereas only MR-Tt expression was detected in glucose-cultured at both the mRNA and the protein level. In addition, the abundance of proteins was examined at different temperatures. At temperatures ranging from 60 to 80 °C, the level of MR-Tt protein was relatively stable, but that of the other proteins was dramatically decreased. The operator-binding complexes were isolated and identified by electrophoretic mobility shift assay-liquid chromatography (EMSA-LC) MS-MS, which suggested that several regulatory proteins, such as GalR and a sensory histidine kinase, participate in the regulation of the operon.

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2009-05-01
2024-03-28
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