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    Computer analysis of the interaction of ribozyme and target RNA
    抗HPV16E7-ribozyme和靶RNA相互作用的计算机分析
    Secondary structure of target RNA impacts on RNA interference efficiency
    目标序列二级结构对RNA干扰效果的影响
    We have analyzed in vitro cleavage reactions of anti-HPV16 E7-ribozyme by computer program. The results demonstrate that secondary structure and energy changes of ribozyme and target RNA affectthe cleavage activity of ribozyme.
    应用计算机分析了抗HPV16E7-ribozyme的体外切割反应结果,发现ribozyme和靶RNA的二级结构及其能量变化能影响ribozyme的切割活性。
    The active ribozyme RNA and the target RNA were generated by in vitro transcription with T7 RNA polymerase.
    以体外转录的PLRV-Ch复制酶基因负链RNA作为底物,与转录的核酶RNA共同保温,以检测核酶对底物的体外切割作用。
    After in vitro transcription, the cleavage reaction was shown to have cut off 79.3% target RNA in 1h.
    经体外转录出核酶RZ1及靶RNA后,体外切割反应1小时,79.3%的靶RNA即被切开。
    Objective To study the in vitro cleavage of mdr1 target RNA by ribozyme and its influencing factors.
    目的 探讨体外核酶对肿瘤多药抗性相关基因mdr1靶RNA分子的切割作用及其影响因素。
    Methods: Rat caspase-3 gene fragment was cloned into T-vector under the control of T7 promoter. 32P-labeled caspase-3 transcript was target-RNA.
    方法:大鼠Caspase-3基因的PCR片段克隆于T载体T7 启动子下游,32P标记的体外转录物作为靶RNA。
    HPV11 E2 mRNA was taken as the target RNA. Ribozyme were designed accounting to the hammerhead structure described by Symons.
    借助计算机软件分析 ,设计出能特异性切割HPV11型 6 4 4ntE2mRNA的核酶 (ribozyme) .
    HPV11 E2 mRNA was taken as the target RNA, ribozyme were designed accounting to the hammerhead structure described by Symons.
    借助计算机软件分析 ,设计出能特异性切割HPV11型 644ntE2mRNA的核酶。
    Methods Rat caspase-3 gene fragment was cloned into the pGEM-T EASY vector under the T7 promoter control. The 32 P-labeled caspase-3 transcript was the target-RNA.
    方法 将大鼠Caspase 3基因的PCR片段克隆于T载体T7启动子下游 ,32 P标记的体外转录物作为靶RNA。
    Results A 198 bp GPI-PLD competitive RNA was constructed and prepared. The competitive RNA could compete well with the target RNA in the RT-PCR reaction.
    结果 :构建和制备了长度为 198bp的GPI PLD竞争性RNA模板 ,该模板与靶模板呈明显的竞争性RT PCR反应 ;
    The 32 P labled caspase 3 transcript being target RNA, cleavage reaction in vitro showed that pM1 GS716 and pM1 GS337 were active, the extent of pM1 GS716 cleavage was 93%.
    3 2 P标记的caspase 3基因片段体外转录物作为靶RNA ,体外切割实验表明 ,pM1 GS716和pM1 GS337均有切割活性 ,其中pM1 GS716的切割效率可达到 93% .
    Conclusion: NHE 1 hammerhead ribozyme can cleave the target RNA specifically, reduce the expression of NHE 1 mRNA, induce intracellular acidosis and consequently prohibit the proliferation of PASMCs.
    结论 :NHE 1特异性锤头状核酶可对NHE 1mRNA进行特异性切割 ,减少其表达 ,从而诱导细胞酸化 ,抑制肺动脉平滑肌细胞增殖
    Guide sequences(GSs) are small RNA molecules that consist of a sequence complementary to a target mRNA and render the target RNA susceptible to degradation by the catalytic RNA subunit of RNase P from Escherichia coli( M1 RNA).
    引导序列GSs(GuideSequences)是能与mRNA互补 ,引导核酶RNaseP催化核心M1RNA对互补区域特异切割的小片段游离RNA。
    GS was designed to the target the mRNA encoding the major DNA polymerase of human cytomegalovirus(HCMV) for degradation. Covalent attachment to the 3′ end of M1 RNA of the guide sequence to DNA polymerase mRNA(M1GS-T7) results in very efficient cleavge of the target RNA in vitro.
    针对人巨细胞病毒HCMV(humancytomegalovirus)DNA聚合酶mRNA序列设计GS ,共价结合到大肠杆菌来源M1RNA中 ,构建成M1GS T7核酶。
    Conclusions The anti-PDGFR- β ribozyme can be expressed stably in HSCs, cleave the target RNA effectively, inhibit HSCs proliferation and collagen synthesis, and induce HSC apoptosis.
    结论 抗PDGFR-β核酶的真核表达载体可在细胞内稳定表达,能有效切割靶RNA,抑制HSC增殖及胶原合成,并诱导其凋亡。
    Objective To investigate whether specific deoxyribozymes aimming at hepatitis C virus(HCV)5′-noncoding region(5′-NCR)have efficient inhibition effect on the target RNA in transgene HepG2.9706 cells.
    目的 观察特异性脱氧核酶 (DRz)在丙型肝炎病毒 (HCV) 5′ 非编码区 (5′ NCR)转基因肝癌细胞株中对靶RNA的抑制活性。
    Methods The fragments of the second exons of pro alpha 1Ⅰand Ⅲcollagen genes were cloned in-to the plasmids pT-Ⅰand pT-Ⅲand labeled with 32 P during transcription to obtain the target RNA.
    方法将含α1Ⅰ型及Ⅲ型前胶原基因第2外显子片段的重组质粒(pT-Ⅰ、pT-Ⅲ),经体外32P标记转录后形成产物靶RNA。
    Method Hammerhead ribozyme against CⅡTA recognizing at 134, 218 and 464 sites (Rz134, Rz218, Rz464 respectively) and CⅡTA target RNA were constructed, then cloned into the pGEM-T vector respectively. The recombinant ribozymes and their CⅡTA target RNA were incu -bated in cell-free conditions.
    方法 设计并克隆针对MHCⅡ类分子转录激活因子(CⅡTA)第134、218、464位点的3 个核酶片段(Rz134、Rz218、Rz464)及其相应的CⅡTA靶基因,分别插入pGEM T载体,进行细胞外切割活性筛选。
    It showed that only Rz464 could cleave target RNA exclusively, then it was cloned into the pIRES2-EGFP vector for intracellular analyses (pIRES2-EGFP-Rz464, pRz464). Stable transfectants of Raji cell by pRz464 were detected for classical MHCⅡ (HLA-DR, -DP, -DQ) expression by flow cytometry, and the mRNA of CⅡTA was detected by RT-PCR.
    将切割作用明显的Rz464 亚克隆入真核表达载体pIRES2 EGFP,成为pIRES2 EGFP Rz464(pRz464),将pRz464稳定转染Raji细胞株,流式细胞术检测Raji细胞表面MHCⅡ类抗原(HLA DR、DP、DQ)的表达,并用逆转录聚合酶链反应分析Raji细胞CⅡTA mRNA的表达。
 

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