LXRα-shRNA真核表达质粒的构建及抑制LXRα表达的有效序列筛选.pdfVIP

1632 生固盔生芏筮查!!!!年!旦筮墼鲞 张琴彭俊1沈薇(重庆医科大学附属第二医院消化内科，重庆400010) 【摘要] 染试剂PalyJelTM将t条质粒转染拿HepG2．2 mRNA和蛋白 效率最佳的质粒。结军三个质粒经测序分析与没汁的序列完全一致．酶切凝胶电泳证实质粒构建成功。三个隧牲均能抑制LXRa 抑制转染细胞中LXRa的表达．其中第一条质粒shRNA-1．XRa!十扰效率最好，从而为脂肪肝的治疗提供F新的方法和材料。 (关键词]肝x受体d；RNA干扰：质粒；掏建 r中圈分类号】R39^3(文献标识码]A[文章编号】1005．9202(2012 ofLXRaand ofeffective toinhibitLXRa Constructionofeukaryoficexpressionplasmid screeulng sequences expression w吐 ZHANG Jnn，SHEN Qin。PENG SecondAffiliated MedicaI ofGastroenterology．theHospital，ChongqingUniversity，Chongqing400010．China Department To tile of thatcaninhibitLXRctmRNAand constructandscreen 【Abstractl0bjecttve plasmidI．XRotgene proteinexpression． wasobtainedfromNCBlwebsite．Threesinail RNA wereselected MethodsHumanLXRdcDNA intefie．ring seqoerlee sequences through online software toRNAi double—strandedDNAwasconnectedwith design accordingdesignprinciples．7thecorresponding pGenesil—I．1plas· mid an(1 andDNA confimledtheeDNA and vector，namelyshRNA，LxRal，shRNA—LxROL2shRNA—LXR∞，Digestionsequencing sequence of transfectedjnto cellswith toassessthetrans— theorientationtheinsert．Threewere HepG2．2．15 pltLsmids PolyJetTM-reagentrespectively feetion underfluorescence andtodeterminethe ofLxRn RT-PeRandWesternblot．ResultsIt efficieney microscope expression gene．wilb DNA was