Cloning and Bioinformatics Analysis of 2-Ketogluconate Transporter Gene from Pseudomonas plecoglossicida
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Abstract:
The aim of this study was to clone the gene kguT encoding 2-ketogluconate transporter from Pseudomonas plecoglossicida JUIM01 and to find its biological information. The degenerate primers were designed on the basis of available sequences of pseudomonads and the physical maps of the 2-ketogluconate utilization operon (kgu operon), and the kguT with the entire nucleotide sequences of 1278 bp was amplified from the strain JUIM01 by TD-PCR. The kguT was predicted to share 87% sequence identity with the corresponding gene encoding 2-ketogluconate transporter (KguT) from Pseudomonas sp. CCOS191, and encode 425 amino acids. The KguT was a hydrophobic transmembrane protein with 12 membrane-spanning domains located in the cell membrane, without signal peptide, and was predicted to share 90% sequence identity with the KguT from Pseudomonas sp. M1. The predicted secondary structure of the KguT contained 75.76% of α helixes, 2.12% of extended strand and 22.12% of random coil. The kguT gene was successfully cloned for the first time from the strain used in industrial production of 2-ketogluconic acid and analyzed by the bioinformatics methods. The results of this study may provide a foundation for further study about the function of 2-ketogluconate transporter from Pseudomonas plecoglossicida JUIM01.
