Molecular Cloning and Bioinformatics Analysis of araC Gene of Vibrio alginolyticus

[Objective] To clone araC gene of Vibrio alginolyticus HY9901 strain, and analyze bioinformatics. [Methods] the whole genome sequence of Vibrio alginolyticus on GenBank was used to design specific primers. According to the principle of PCR amplification sequence, the target gene araC was amplified, and then the sequence was further analyzed by bioinformatics method to establish the phylogenetic tree of araC gene and its corresponding subunit three-dimensional structure model. [Results] Sequence analysis revealed araC gene is 711 bp and encodes a putative protein of 236 amino acids. The predicted molecular mass of AraCwas 26.92 ku. Using Signal P 4.0 and TMHMM Server 2.0 software for analysis, it was predicted that the AraC protein did not contain a signal peptide or a transmembranous region. The AraC protein had two cAMP and cGMP dependent protein kinase phosphorylation site, five protein kinase C phosphorylation sites, three casein kinase II phosphorylation sites, one prenyl group binding site (CAAX box) and five microbodies C-terminal targeting signal. The predicted results of protein subcellular localization showed that AraC was located in the mitochondria, nucleus and cytoplasm. Its protein is unstable and hydrophilic. The AraC protein is a transcriptional regulatory protein which belongs to HTH_18 superfamily. According to the prediction, secondary structure: a-helix (Alpha helix) accounted for 52.12%, Random coil (31.78%), Extended strand (11.02%), b-fold (Beta turn) accounted for 5.08%. V. alginolyticus, Vibrio parahaemolyticus and Vibrio palustris were clustered together, which implies that the genetic relationship between these three species was the closest.