一株肠炎沙门氏菌鞭毛基因FliC的克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.12.009

Abstract

Abstract:

This study was aimed to clone the flagellin FliC gene of Salmonella Enteritidis and analyze its genetic structure with bioinformatics.The genomic DNA of bacterium was extracted as a template,according to the published sequence of FliC gene in GenBank,and one pair of specific PCR primers was designed.FliC gene was cloned by PCR and linked into the cloning vector to be sequenced.The nucleotide sequence,amino acid sequence,homology and phylogenetic tree were analyzed by the bioinformatics softwares.The physicochemical properties,hydrophilicity,signal peptides,transmembrane domains,glycosylation sites,B cell epitopes,secondary structure and tertiary structure of the encoding protein were predicted by DNAStar software and online servers.The results showed that the gene of 1 518 bp was successfully cloned and encoded 505 amino acids.Homology analysis showed that FliC gene was relatively conserved,and this species had high homology with E.coli genus.The chemical formula of this protein was C₂₂₅₄H₃₇₀₁N₆₅₇O₈₀₃S₄,and the theoretical molecular mass was 52.981 ku.The theoretical isoelectric point was 4.91,and the instability index was 16.86,which was a stable hydrophilic protein.Structural analysis result showed that neither a signal peptide nor a transmembrane domain was found,however,a total of 8 N-linked glycosylation sites,13 O-linked glycosylation sites,60 phosphorylation sites,26 linear B cell piptopes and 5 T cell binding sites were predicted.Secondary structure analysis showed that α-helix,β-turn,extended chain and random coil accounted for 43.76%,3.76%,20.99% and 31.49%,respectively.In this study,FliC gene was cloned and analyzed successfully.The results laid a theoretical foundation for the further study of the role of flagella in the pathogenesis and the development of genetic engineering vaccines.

Key words: Salmonella Enteritidis; flagella; FliC gene; bioinformatics analysis

CLC Number:

S855.1⁺2

CHENG Xiangchao, TIAN Wenjing, ZHANG Mengke, MAO Fuchao, LIAO Chengshui. Cloning and Bioinformatics Analysis of a Flagellar FliC Gene in Salmonella Enteritidis[J]. , 2018, 45(12): 3379-3386.

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Cloning and Bioinformatics Analysis of a Flagellar FliC Gene in Salmonella Enteritidis

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