人血管内皮生长子受体-3胞外1-3区基因的表达.pdfVIP

人血管内皮生长因子受体一3胞外卜3区基因的表达 摘要 目的 1血管内皮生长因子受体．3基因的原核表达载体构建 2．血管内皮生长因子受体一3基因的原核融合表达 方法 以本实验室构建的克隆质粒为模板，设计合适引物，在上游引物中加入酶切位点和起 始密码，在下游引物中加入终止密码和酶切位点，把上述VEGFR3扩增产物经切胶回收纯 肠杆菌DH5 a，筛选出阳性菌株，温控诱导表达，超声破菌，对包涵体进行洗涤纯化．以 并转化大肠杆菌DH5 结果 Ni2+_NTA树脂柱纯化后获得VEGFR3融合蛋白纯品。 结论 为进一步的研究奠定了基础。 关键词 人血管内皮生长因子受体一3，蛋白表达，融合 ofHumanVascularEndothelial Expression GrowthFactor 3ExtracellularDomam1-3 Receptor Master Degree candidate：Caojing Director：Professor WangBinquan Major：Otolaryngology oftheFirst ofShanxiMedical Department Hospital University Abstract objective： 1：Toconstructthe vectorofHumanVEGFR3 prokaryoticexpression 2：TO thefusion ofHumanVEGFR3． express protein Method： We 8T-VEGFR3whichconstructed byour designedproperprimer．ThepMDI previously WaSusedaS and withstream EcoRIsiteand laboratory templateamplifiedup primercontaining stream BamHI site．Aftercollectionand subclonedthis down primercontaining purification，we was PCR into andtransformedEcoliDH5Ⅱ．The bacteriaselected productpBV220 positive andinducedat42℃．Thebacteriawasbrokenultrasonication．Theinclusion body by containing VEGFR3was andwashed．Alsoweconstructed of plasmid targetprotein separated expression VEGFR3fusion