人脑源性神经营养因子成熟肽基因克隆及表达研究.pdfVIP

人脑源性神经营养因子成熟肽基因克隆及表达 张虎1杨吉成盛伟华缪党诚’李丽娥王金志国艳艳落掌锋 苏州大学医学院基础医学系细胞与分子生物攀教研室，澄苏，苏州215007 摘要：目的：构建表达人脑源性神经营养因子的基因工糕菌。方法：人工合成一对含 EcoRI和BamHI限制性酶切位点的特异引物，以健康成人末梢囱细胞基因缀DNA作为模板。 DH5a经热诱导表达。结果：经DNA测序和限制性酶切确证燕组入载体片段为羁酌基因的 核营酸寄烈，并．鑫翡表达蛋叁。结论：本研究或功遥克隆毒hBDNT基因缡码亭酬并在大瑟 杆菌中获得稳定表达，为今后的生物学活性和神缀系统的基因治疗奠定了基础 基因表达。 关键调：齄源性神经营养困子；PCR方法；A-T克隆 constructthe bacteriumforhuman ．Abstract：Objective：T0geneticallyengineered of brai_．n-derived factorMethods：A the site specific including neurotrophic pair primers cleavage of endonucleaseEcoRlandBamHIhasbeen the DNA restriction synthesized．from genomicBy PCRtheBDNF was fromthe DNAisolatedfrom using genesequencedirectlyampLifiedgenomic of wasthenclonedintO vector．The thehuman ffazment Pucm-T leucocytes。．the resultinggene insert fromtherecombinant was withtinearied fragment pUCm，■BDNFdirectionaltyligated PBV220．DH5awastransformedwiththe recombinantPBV220．BDNFandinduced expression by wasidentifiedasthe laB王)NF heatResult：TheclonedGene sequence b￥D慕A encoding and restriction revealedthecloned sequencingsequencing anaylsis．SDS—B气GE geneexpressed the human is Condusion：Theclone BDNFobtained successfully containingsequenceencoding in