Cloning And Purification of a Putative Repressor Protein from Arthrobacter nicotinovorans Pao1

Besides the well-characterized pathway for nicotine degradation (Brandsch, 2006), pAO1 carries a gene cluster of a hypothetical pathway for carbohydrate utilization. This cluster consists of ORFs of a transcriptional regulator, of a sugar ABC-transporter and of several putative dehydrogenases and oxidoreductases. Previously, we established that the pAO1 orf39 gene encodes an aldehyde-dehydrogenase (Mihasan, 2010) and orf40 encodes an sugar dehydrogenase. Here we focus on further characterization the ORF32 protein and elucidation of its possible role in the cell. By cloning the gene in the plasmid vector pH6EX3, we were able to express it as a recombinant His-tagged protein and to easily purify it to homogenity.