摘要
[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.
[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.
基金
Supported by Natural Science Foundation of Guangdong Province(2017A030313174)
"Sail of the Sea-Starting Plan" of Guangdong Ocean University(qhjhzr201813)
Innovation and Entrepreneurship Training Program for College Students(No.CXXL2019041)
