Cloning and Bioinformatics Analysis of tye A Gene of Vibrio alginolyticus

[Objectives]This study aimed to clone the tye A gene of Vibrio alginolyticus HY9901 strain and analyze its sequence by bioinformatics.[Methods] By referring to the entire genome sequence of Ⅴ.alginolyticus on Gen Bank,specific primers were designed.According to the principle of PCR amplification,the target gene tye A was amplified.By means of bioinformatics,the sequence of tye A was further analyzed,and the phylogenetic tree of tye A genes of Vibrio spp.and the corresponding subunit three-dimensional structure models were constructed.[Results] The length of the tye A gene of Ⅴ.alginolyticus strain HY9901 is 285 bp,and its theoretical molecular weight is 10.98 kD.According to prediction,there is no signal peptide or transmembrane region at the N-terminus of the sequence,and the amino acid sequence contains two casein kinase Ⅱ phosphorylation sites.The results of protein subcellular localization prediction show that the Tye A protein is located in the cell membrane.The protein is unstable and non-hydrophilic.The tertiary structure of Tye A protein of Ⅴ.alginolyticus is similar to that of Yersinia sp.According to prediction,Tye A has a major functional domain Pfam.In terms of secondary structure,alpha helix,random coil,extended strand and beta turn account for 85.11%,7.45%,4.26% and 3.19%,respectively.The homology of Tye A between Ⅴ.alginolyticus and Vibrio parahaemolyticus is up to 83%,so they are classified into one cluster.[Conclusions]This study will help to further understand the regulation mechanism of type Ⅲ secretion system in Ⅴ.alginolyticus.

Inhibitory Effects of Prunus mume,Coptis chinensis,and Crataegus pinnatifida on Vibrio parahaemolyticus and Its Biofilm in Vitro

In order to explore the inhibitory effects of Prunus mume,Crataegus pinnatifida and Coptis chinensis on Vibrio parahaemolyticus and its biofilm in vitro,the agar diffusion method was applied. These three Chinese herbal medicines had different inhibitory effects on V. parahaemolyticus. The inhibition zone of C. pinnatifida to V. parahaemolyticus was( 15. 25 ± 0. 53) mm,and the minimum inhibitory concentration( MIC) and minimum bactericidal concentration( MBC) of C. pinnatifida on V. parahaemolyticus were both 31. 25 mg/m L; the inhibition zone of C. chinensis to V. parahaemolyticus was( 18. 08 ± 0. 10) mm,and the MIC and MBC of C. chinensis on V. parahaemolyticus were both15. 63 mg/m L; the inhibition zone of P. mume to V. parahaemolyticus was( 28. 99 ± 0. 47) mm,and the MIC and MBC of P. mume on V. parahaemolyticus were both 7. 81 mg/m L. The effects of three traditional Chinese medicines on the biofilm formation of V. parahaemolyticus were tested by MTT colorimetric method using methylthiazolyl tetrazolium( MTT). P. mume,C. pinnatifida and C. chinensis have significant inhibitory effects on V. parahaemolyticus biofilm and their MIC are 7. 81 mg/m L,3. 125 mg/m L,and 62. 5 mg/m L,respectively( P < 0. 01).The experimental results are expected to provide certain references for the development of new fishery drugs.