Survivin反义RNA%2fHSP70双基因表达载体的构建及鉴定.pdf

中 文摘要 肿瘤细胞增殖、诱导肿瘤细胞凋亡以及激发机体有效的特异性抗肿瘤免疫应答提供重要 的实验材料。方法应用RT—PCR从Jurkat细胞中获得SurvivineDNA片段，反向插入 是否成功；应用RT—PCR从HepG2细胞中获得的HSP70eDNA片段．定向插入到 是否正确。结果经酶切和测序鉴定证明Survivin反义tLNA表达载体已成功构建；经 菌落PCR、限制性酶切和测序鉴定证明HSP70表达载体已成功构建。结论本实验已 成功构建了Survivin反义RN～HSP70双基因表达载体，为进一步研究提供了实验基础。 主题词 SurvivinHSP70 RT—PCR反义RNA Constructionandindentification ofSurvivin antisense RNA／HSP70 vector double—。geneexpression ABSTRACT AnSurvivtnantisence Objective RNA／HSP70 vectorwas double—geneexprer商on constructedtO materialforthefurther provideimportantexperimental research。wIlichwas usedtOtransfectthe U．nTlorcells，inhibit ofttnnor oftumor proliferationcells，induce apoptosls ceils andstimulatethe anti—t1．mlorinl／nuYle specific respome AeDNA effectively．Methods survivin fragmentof obtainedRT—PCRwasinserted by intoa vector珑酬 plasmid inthereverse pIRES2一DaRed2direction．SurvivinantisenseKNAwas cordirmedre． using smction andDNA enzymedige!sion HSPT0eDNA sequencing．A whichWaSob- fragmem talned fromthe cell HepG2 RT—PCRwasime．rtedinto