expansin和expansinlike蛋白及其作用机制的研究word格式论文.docxVIP

expansin-likeproteinTre1fromfungusTrichodermareesei.WeclonedtheTre1cDNA sequence(1383bp)includingstartcodonandterrminationcodonbyRACE-PCR,andTre1genomicDNAsequence(1725bp)includinganintronandpolyAbyinversePCR.TheresultofgeneanalysisandalignmentshowedthatTre1wasanewgene,andtheTre1proteinsequenceincludedacompleteGlycosylhydrolasefamily30conserveddomain.ButtherecombinantexpressionofTre1showedthatnoactivesolubleproteincouldbedetectedbothinE.coliandP.pastoris.TheactivityofrecombinantTre1proteinmaybedependson thepost-translationalmodificationtoo.Ontheotherhand,consideringweclonedTre1 fromtheATCCstandardstrainwhichmightbedifferentingenecodonsfromtheindustrystrain,wehopetogettheactiveTre1proteinmutantbymolecularrevolution.Keywords:expansin,swellingfactor,hydrolyticenzymes,synergism,expansin-likeprotein,heterogenousexpression符号说明BSAbovine serum albumin牛血清白蛋白CBDcellulose binding domain纤维素结合域cDNAcomplementaryDNA互补DNA CTABcetyltrimethylammoniumbromide十六烷基三乙基溴化铵DNSdeinitrosalicylic acid二硝基水杨酸E.ColiEscherichiacoli大肠杆菌EGaseendo-glucanase葡聚糖内切酶Hishistidine组氨酸IPTGIsopropylβ-D-1-Thiogalactopyranoside异丙基-β-D-硫代半乳糖苷LB mediumLuria-BertaniMediumLB培养基MLGmixed-linkage-(1-3),(1-4)-glucans混合键葡聚糖NCBInationalcenterforbiochemstryimformation美国国立生物信息中心ODOptical densit光密度PAGEpolyacrylamidegelelectrophoresis胶体电泳PCRpolymerasechainreaction/polymerasecyclicreaction多聚酶链式反应/体外扩增PEGpolyethyleneglycol聚乙二醇pETProtein ExpressionSystems蛋白表达系统PMSFPhenylmethanesulfonylfluoride苯甲基磺酰氟PVDFmembranepolyvinylidenedifluoride membranePVDF膜/聚偏二氟乙烯膜SDSSodiumdodecyl sulfate十二烷基硫酸钠TEMEDN,N,N’,N’-tetramethylethylenediamineN,N,N’,N’-四甲基乙二胺XETxyloglucanendotransglycosylase木葡聚糖内转移酶XGxyloglucan木葡聚糖学位论文独创性声明本人郑重声明：1、坚持以“求实、创新”的科学精神从事研究工作。2、本论文是我个人在导师指导下进行的研究工作和取得的研究成果。3、本论文中除引文外，所有实验、数据和有关材料均是真实的。4、本论文中除引文和致谢的内容外，不包含其他人或其它机构已经发表或撰写过的研究成果。5、其他同志对本研究所做的贡献均已在论文中作了声明并表示了谢意。研究生签名：日期：学位论文使用授权声明本人完全了解南京师范大学有关保留、使用学位论文的规定，学校有权保留学位论文并向国家主管部门或其指定机构送交论文的电子版和纸质版；有权将学位论文用于非赢利目的的少量复制并允许论文进入学校图书馆被查阅；有权将学位论文的内容编入有关数据库进行检索；有权将学位论文的标题和摘要汇编出版。保密的学