Cloning and Bioinformatics Analysis of pepck Gene in Vibrio alginolyticus

[Objectives] To clone the pepck gene of Vibrio alginolyticus strain HY9901 and analyze its sequence by bioinformatics. [Methods] According to the complete gene sequence of V. alginolyticus on GenBank, specific primers were designed to amplify the target gene pepck by PCR. The sequence of the pepck gene was analyzed using bioinformatics. The phylogenic tree of pepck gene and the corresponding single-subunit three-dimensional structure were constructed. [Results] The pepck gene of V. alginolyticus strain HY9901 has a full length of 1 629 bp, with theoretical molecular weight of 60.12 kD. The prediction results show that there is no signal peptide or transmembrane region at the N-terminus of the sequence, the amino acid sequence contains 11 phosphorylation sites of casein kinase II. The prediction results of protein subcellular localization indicate that PEPEK protein is localized in the cytoplasm. The protein is stable and hydrophobic. The tertiary structure of the PEPCK protein of V. alginolyticus is similar to that of Vibrio parahaemolyticus. It is predicted that PEPCK has a major functional domain PEPCK_ATP. In the secondary structure, alpha helix, random coil, and extended strand accounted for 21.96%, 52.03% and 26.01%, respectively. The PEPCK homology between V. alginolyticus and Vibrio diabolicus is as high as 99%. [Conclusions] This study lays the foundation for further understanding the function of pepck gene in V. alginolyticus.