广东海洋大学学报2016，Vol.36Issue(4)：30-36,7.DOI:10.3969/j.issn.1673-9159.2016.04.006
溶藻弧菌III型分泌系统vscP基因的克隆与生物信息学分析
Molecular Cloning and Bioinformatics Analysis of Yop Secretion Protein P (VscP) fromVibrio alginolyticus Strain HY9901
摘要
Abstract
Primers for PCRcloning were designed according to the whole genome sequence ofVibrio alginolyticus published inGenBank. The type 3 secretion system (T3SS) Yop secretion protein P (VscP) gene ofV. alginolyticusstrain HY9901 was amplified by PCR and cloned into pMD 18-T vector. Sequence analysis reveals thatthe ORF of VscP gene (GenBank Accession: FR780678)is 1 206bpand eneodes a putative protein of401 amino acids.The predicted molecular weight (MW) of VscP was44.4 ku with an estimated pI of 5.72.Using SignalServer 4.1and TMHMM Server 2.0 software, and itwas predieted that theVscP protein did not contain a signal peptide or a transmembranous region.TheVscP protein had threeN-glycosylation sites, two cAMP and cGMP dependent protein kinase phosphorylation site,five protein kinase C phosphorylation sites, nine casein kinase II phosphorylation sites, two N-myristoylation sites,one amidation site and three loci microbody C-terminal targeting signal predieted by SoftBerry-Psite software. Thisprotein shows high genetic relationship withVibrio parahaemolyticus via a molecular phylogenetic tree constructed by MEGA6.0software. A model of VscP subunitsapplication of SWISS-MODELwork-space is built andits three-dimensional structureis found to havesimilar configurationwithflagellar hook-length control protein FliK.关键词
溶藻弧菌/III型分泌系统/基因克隆/生物信息学分析/vscPKey words
Vibrio alginolyticus/type 3 secretion system/gene cloning/bioinformaticsanalysis/vscP分类
生物科学引用本文复制引用
李静,庞欢瑛,简纪常,鲁义善,汤菊芬,吴灶和..溶藻弧菌III型分泌系统vscP基因的克隆与生物信息学分析[J].广东海洋大学学报,2016,36(4):30-36,7.基金项目
国家自然科学基金（31402344）；海洋经济创新发展区域示范专项 （）
