翘嘴红鲌热休克蛋白60的基因克隆及其表达特征分 析（英文）.pdfVIP

第35卷 第 2期 浙江海洋学院学报(自然科学版) V01．35 No．2 2016年3月 JournalofZhejiangOceanUniversity(NaturalScience) Mar．，2016 文章编号：1008—830X(2016)02—0105—08 M olecularCloning，Characterization andExpressionAnalysisof HeatShockProtein60inErythroculterilishaeformis DAIDong，YANG Yu-hong (CollegeofAnimalScienceandTechnology，NortheastAgriculturalUniversity，Harbin 150030，China) Abstract：Inthepresentstudy，thecDNAofheatshockprotein60fromErythroculterilishaeformis(EiH— SP60)wasclonedbythecombinationofhomoloyg cloningandrapidamplificationofcDNAends(RACE)ap— preaches．Thefull—lengthcDNAofEiHSP60wasof2323bp，includinganopenreadingframe(ORF)ofl728 bpencodingapolypeptideof575aminoacidswithpredictedmolecularweightof61．26kDaandtheoretical isoelectricpointof5．27．BLAST analysisrevealed thatEiHSP60 shared high similaritywith otherknown HSP60s．andoneconservedChaperonin60signaturewasalSOidentifiedinEiHSP60，which indicatedthat EiHSP60shouldbeamemberoftheHSP60family．ThemRNA ofEiHSP60wasconstitutivelyexpressedinall testedtissuesofuntreatedtopmouthculter，includingbrain，muscle，kidney，liver，skin，spleen，heart，gilland intestinewiththehighestexpressionlevelintheliver．Fluorescentreal—timequantitativeRT—PCR wasusedto examinetheexpressionoftheEiHSP60geneintopmouthculterafterthechallengewithLPS．A cleartime-de- pendentexpression pattern ofEiHSP60wasfoundaftertheLPS challenge。and theup—regulatedmRNA ex— press10nofEiHSP60intopmouthculterafterLPSchallengeindicatesthattheHSP60geneiSinducibleandin- volvedinimmuneresponses．TheseresultssuggestthatEiHSP60playsanimportantroleinimmuneresponses intopmouthcuher． Keywords：Erythroculterilishaeformis；heatshockprotein60；genecloning；characterizationanalysis CLCnumber：S917．4 Documentcode：A 翘嘴红鲔热休克蛋白6O的基因克隆及其表达特征分析 戴 东，杨雨虹