Molecular Cloning and Bioinformatics Analysis of cyaA Gene of Vibrio alginolyticus Strain HY9901.

[Objectives] This study was conducted to explore the biological functions of cyaA gene of Vibrio alginolyticus. [Methods] With DNA of V. alginolyticus HY 9901 as a template, primers were designed according to the sequence of cyaA gene, and the cyaA gene was amplified by PCR. Bioinformatics analysis was performed. [Results] The cyaA gene of V. alginolyticus HY9901 was 2 529 bp in size, and encoded 842 amino acids. The molecular structure of CyaA protein was C_{4 358}H_{6 745}N_{1 171}O_{1 286}S₃₅. Its theoretical molecular weight was 97. 241 67 kDa and the theoretical pi value was 5. 56. It had no signal peptide and transmembrane domain. CyaA protein had three N-terminal glycosylation sites, one cAMP and cGMP-dependent protein kinase phosphorylation site, nine protein kinase C phospho-rylation sites, nine casein kinase II phosphorylation sites, one tyrosine kinase phosphorylation site, seven N-terminal myristoylation sites, one pentenyl binding site and ten microbody C-terminal localization signal sites. Subcellular localization prediction showed that CyaA protein was mainly located in the nucleus and cytoplasm. Through multi-sequence alignment and phylogenetic tree construction, it was concluded that V. alginolyticus had high CyaA homology with other Vibrio species. cyaA of V. alginolyticus was clustered with Vibrio fluminensis and Vibrio marinisedimini, and they were closely related. The secondary structure of CyaA protein consisted of a-helixes (43.11%), random coils (38.00%) and extended strands (14.49%). In protein network interaction, it was found that the proteins adjacent to CyaA protein were Crp-2, CpdA, Crr, PtsG-2, ANP67209.1, Crp-1, PykF, Pyk, RelA and Ndk. [Conclusions] This study provides a new idea for formulating strategies for the prevention and control of vibriosis. [ABSTRACT FROM AUTHOR]