留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

哈维氏弧菌FlaA基因的克隆、序列分析及真核表达质粒的构建

庄轩 覃映雪 苏永全 王军 丁少雄

庄轩, 覃映雪, 苏永全, 王军, 丁少雄. 哈维氏弧菌FlaA基因的克隆、序列分析及真核表达质粒的构建[J]. 海洋学报, 2007, 29(6): 74-79.
引用本文: 庄轩, 覃映雪, 苏永全, 王军, 丁少雄. 哈维氏弧菌FlaA基因的克隆、序列分析及真核表达质粒的构建[J]. 海洋学报, 2007, 29(6): 74-79.
ZHUANG Xuan, QIN Ying-xue, SU Yong-quan, WANG Jun, DING Shao-xiong. Cloning and sequencing of FlaA gene of Vibrio harveyi and construction of its eukaryotic expression recombinant plasmid[J]. Haiyang Xuebao, 2007, 29(6): 74-79.
Citation: ZHUANG Xuan, QIN Ying-xue, SU Yong-quan, WANG Jun, DING Shao-xiong. Cloning and sequencing of FlaA gene of Vibrio harveyi and construction of its eukaryotic expression recombinant plasmid[J]. Haiyang Xuebao, 2007, 29(6): 74-79.

哈维氏弧菌FlaA基因的克隆、序列分析及真核表达质粒的构建

基金项目: 国家863项目(2003AA603011);福建省重大科技项目(2002N009)

Cloning and sequencing of FlaA gene of Vibrio harveyi and construction of its eukaryotic expression recombinant plasmid

  • 摘要: 哈维氏弧菌是水产病害中常见的致病菌.参照基因库上登录的弧菌鞭毛丝蛋白FlaA基因序列设计简并引物,PCR扩增哈维氏弧菌TS-628株的FlaA全基因,并克隆到pMD 18-T载体中测序,经序列分析该基因全长1 140 bp,编码379个氨基酸.与基因库中其他弧菌的同源基因序列比较显示,哈氏弧菌FlaA基因与霍乱弧菌FlaA基因的同源性最高(79.5%),该基因编码的多肽缺乏半胱氨酸,并在N-,C-两端的氨基酸序列较为保守,中间区域的变异较大.对该蛋白的氨基酸组成及空间结构进行了分析和预测,并推测该蛋白质的平均分子量为40.6 kDa.在该基因末端加上一段编码Flag短肽的核苷酸序列后克隆到真核表达载体pcDNA3.1(+),获得带哈氏弧菌鞭毛丝蛋白FlaA基因的真核表达重组质粒pcDNA-FlaA-tag,为其DNA疫苗的进一步研究奠定了基础.
  • COPLAND J W,GREY D L.Management of wild and cultured sea bass/barramundi(Lates calcarifer Bloch)[A].Proceeding of an International Workshop Help at Darw in N T,Australia[C].ACIAR Proceedings,1987.20.
    AOKI T,EGUSA S.Drug sensitivity of Aeromonas liquefaciens isolated from freshwater fishes[J].Bull Japan Soc Fish,1971,37(3):176-185.
    FUDA H,SOYANO K,YAMAZAKI F,et al.Serum immunoglobulin(IgM) during early development of masu salmon(Oncorhychus masou)[J].Comp Biochem and Physiol,1991,99A:637-643.
    相建海.海水养殖生物病害发生与控制[M].北京:海洋出版社,2001.
    FYNAN E F,WEBSTER R G,FULLER D H,et al.DNA vaccines:A novel approach to immunization[J].Int J Immunopharmac,1995,17(2):79-83.
    HEPPELL J,DAVIS H L.Application of DNA vaccine technology to aquaculture[J].Advanced Drug Delivery ReviewS,2000,(43):29-43.
    HEPPELL J,LORENZEN N,ARMSTRONG N K,et al.Development of DNA vaccines for fish:vector design,intramuscular injection and antigen expression using viral hemorrhagic septicemia virus genes as model[J].Fish Shellfish Immunol,1998,8:271-287.
    周化民,苏永全,王军,等.副溶血弧菌(Vibrio parahaemolyticus)FlaI基因的合成及其克隆与鉴定[J].自然科学进展,2002,12(1):101-103.
    覃映雪,苏永全,周化民,等.副溶血弧菌极鞭毛蛋白FlaB基因的克隆与序列分析[J].农业生物技术学报,2003,11(6):656-657.
    陈奖励.水产微生物学[M].北京:农业出版社,1993.474-478.
    DAS M,CHOPRA A K,WOOD T,et al.Cloning,sequencing and expression of the flagellin core protein and other genes encoding structural proteins of the Vibrio cholerae flagellum[J].FEMS Microbiology letters,1998,165:239-246.
    KIM Y K,MCCARTER L L.Analysis of the polar flagellar gene system of Vibrio parahaemolyticus[J].J Bacteriol,2000,182:3 693-3 704.
    MCGEE K,HORSTEDT P,Milton D L.Identification and characterization of additional flagellin genes from Vibrio anguillarum[J].J Bacteriol,1996,5 188-5 198.
    覃映雪,池信才,苏永全,等.网箱养殖石斑鱼溃疡病病原的研究[J].水产学报,2004,28(3):297-302.
    SAMBROOK J,FITCH E,MANIATIS T.Molecular Cloning:A Laboratory Manual.2nd ed.[M].New York:Cold Spring Harbor Laboratory Press,1989.
    HOMMA M,FUJITA H,YAMAGUCHI S,et al.Regions of Salmonella typhimurium flagellin essential for its polymerization and excretionp[J].J Bacteriol,1987,169:291-296.
    KUWAJIMA G,KAWAGISHI I,HOMMA M,et al.Export of an N-terminal fragment of Escherichia coli flagellin by a flagellum-specific pathway[J].Proc Natl Acad Sci USA,1989,86:4 953-4 957.
    WINSTANLEY C,MORGAN A W,PICKUP R W,et al.Molecular cloning of two Pseudomonas flagellin genes and basal body structural genes[J].Microbiology,1994,140:2 019-2 031.
    YANG E C H,SCHRANK G D,FREEMAN B A.Purification of Flagellar Cores of Vibrio cholerae[J].J Bacteriol,1977,1 121-1 128.
    TOTTEN P A,LORY S.Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK[J].J Bacteriol,1990,172:7 188-7 199.
    MILTON D L,O'TOOLE R,HORSTEDT P,et al.Flagellin A is essential for the virulence of Vibrio anguillarum[J].J Bacteriol 1996,178(5):1 310-1 319.
    覃映雪,王军,王世峰,苏永全,庄轩.重组质粒pcElaA在青石斑鱼肌肉中的表达研究.高技术通讯(待刊).
  • 加载中
计量
  • 文章访问数:  946
  • HTML全文浏览量:  13
  • PDF下载量:  1116
  • 被引次数: 0
出版历程
  • 收稿日期:  2006-05-10
  • 修回日期:  2006-10-23

目录

    /

    返回文章
    返回