Cloning And Purification of a Putative Repressor Protein from Arthrobacter nicotinovorans Pao1
Poster Jul 24, 2013
Marius Mihasan, Marius Stefan, Hritcu Lucian, Vlad Artenie
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.
Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
Concern about the safety of food products has increased dramatically with intentionally and non-intentionally added substances (NIAS) used in packaging being of particular interest. APGC and high resolution MS together with a workflow driven process was used to identify unknown compounds in packaging.READ MORE