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target rna
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  靶rna
    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。
短句来源
    METHODS Three single ribozymes against hepatitis B virus core gene were designed with computer, then transctiptional vector of three singleribozymes were constructed, HBV protein influence on the cleavage activity of single ribozyme on target RNA observed, and HBV protein interaction with target RNA further analysed.
    方法 利用计算机辅助设计针对乙型肝炎病毒 C基因的三个单一核酶构建 Rz1核酶自剪切转录载体 (p GEMRz1) ,观察单一核酶 (Rz1)对靶 RNA的切割作用及乙型肝炎病毒蛋白对核酶剪切作用的影响 .
短句来源
    RESULTS Single antiviral ribozyme(Rz1) could cleave its target RNA efficiently, HBV proteins had no effects on ribozyme mediated cleavage on HBV. Further results indicated that HBV proteins did not interact with target RNA in our experiment.
    结果 构建的核酶自剪切转录载体体外转录后 ,在顺式核酶发生自剪切后可将目的核酶正确地释放出来 ,计算机设计的单一核酶体外可剪切靶 RNA; 乙型肝炎病毒蛋白对核酶剪切的无明显的抑制或增强作用 ,进一步的结果未观察到乙型肝炎病毒蛋白和靶 RNA的相互作用 .
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  “target rna”译为未确定词的双语例句
    Result Under the adopted conditions the three deoxyribozymes efficiently cleaved the target RNA in vitro and the cleavage activity of DRz3 was increased with the increase of Mg~(2+)concentration.
    随着 Mg~(2+)浓度的增加,DRz3的切割 活性增强。
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  target rna
Both ribozymes were first assayed on a synthetic 16 bases target RNA and found to catalytically and efficiently cleave the substrate in a sequence specific way.
      
In this chapter we describe methodologies to first analyze the ribozyme catalytic activity in vitro by studying cleavage of target RNA in vitro.
      
The behavior of class II mutants requires that exogenous 5FU is specifically excluded from the site of synthesis of the target RNA involved in petite mutagenesis, while 5FC has access to it.
      
In several systems, silencing has been found to spread from the dsRNA inducer sequence into upstream or downstream regions of the target RNA, a phenomenon termed transitive silencing.
      
The target RNA species may be the products of transgenes, endogenous plant genes or viral RNAs.
      
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AIM To detect HBV protein influence on the cleavage activity of ribozyme. METHODS Three single ribozymes against hepatitis B virus core gene were designed with computer, then transctiptional vector of three singleribozymes were constructed, HBV protein influence on the cleavage activity of single ribozyme on target RNA observed, and HBV protein interaction with target RNA further analysed. RESULTS Single antiviral ribozyme(Rz1) could cleave its target RNA efficiently, HBV proteins...

AIM To detect HBV protein influence on the cleavage activity of ribozyme. METHODS Three single ribozymes against hepatitis B virus core gene were designed with computer, then transctiptional vector of three singleribozymes were constructed, HBV protein influence on the cleavage activity of single ribozyme on target RNA observed, and HBV protein interaction with target RNA further analysed. RESULTS Single antiviral ribozyme(Rz1) could cleave its target RNA efficiently, HBV proteins had no effects on ribozyme mediated cleavage on HBV. Further results indicated that HBV proteins did not interact with target RNA in our experiment. CONCLUSION HBV proteins has no effects on ribozyme mediated cleavage on HBV.

目的 探讨乙型肝炎病毒蛋白对核酶剪切作用的影响 .方法 利用计算机辅助设计针对乙型肝炎病毒 C基因的三个单一核酶构建 Rz1核酶自剪切转录载体 (p GEMRz1) ,观察单一核酶 (Rz1)对靶 RNA的切割作用及乙型肝炎病毒蛋白对核酶剪切作用的影响 .结果 构建的核酶自剪切转录载体体外转录后 ,在顺式核酶发生自剪切后可将目的核酶正确地释放出来 ,计算机设计的单一核酶体外可剪切靶 RNA;乙型肝炎病毒蛋白对核酶剪切的无明显的抑制或增强作用 ,进一步的结果未观察到乙型肝炎病毒蛋白和靶 RNA的相互作用 .结论 体外转录的核酶可剪切乙肝病毒靶 RNA,在本研究条件下未观察到乙肝病毒蛋白对核酶切割作用的影响 ,从而确保核酶在细胞内能有效发挥作用 .

Objectives To study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor β subunit (PDGFR- β) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs. Methods Expression vector of anti-PDGFR- p ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR-β , α -smooth muscle actin (α -SMA), and type I and type in collagen was...

Objectives To study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor β subunit (PDGFR- β) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs. Methods Expression vector of anti-PDGFR- p ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR-β , α -smooth muscle actin (α -SMA), and type I and type in collagen was detected by means of northern blot, Western blot and immunocytochemical staining respectively. The cell proliferation was determined with MTT colori-metric assay. The cell apoptosis was demonstrated with flow cytometry. acridine orange fluorescence vital staining and transmission electron microscopy. Results The expression of PDGFR- BBBBB at mRNA and protein level was markedly reduced in ribozyme-transfected HSCs only 43%-51% of that in control cells (t ≥ 3.957, P < 0.05), and α-SMA expression level, type Ⅰ and type Ⅱ collagen synthesis ability were also reduced (t ≥ 6.790, P < 0.01). The proliferation of ribozyme-transfected HSCs was significantly decreased (t ≥3.858, P < 0.05), and the proliferation response to PDGF BB was markedly inhibited. However the apoptotic rate was significantly increased in ribozyme-transfected HSCs ( x2 ≥ 14.157, P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy. 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. The results suggest that inhibiting PDGFR- β expression in HSCs may be a new therapy for liver fibrosis.

目的 研究抗血小板衍生生长因子受体β亚单位(PDGFR-β)核酶在肝星状细胞(HSC)内的切割活性及其对HSC生物学特性的影响。 方法 构建抗PDGFR-β核酶的真核表达载体,将其转染入HSC-T6细胞,G418筛选出阳性细胞克隆;分别用northern blot、western blot和免疫细胞化学检测PDGFR-β表达,用MTT法检测细胞增殖,免疫细胞化学检测α-F滑肌肌动蛋白(α-sMA)和Ⅰ、Ⅲ型胶原表达,用流式细胞仪、吖啶噔荧光染色和电镜分析细胞凋亡。 结果 转染核酶的HSC的PDGFR-β在mRNA和蛋白水平的表达量均显著降低,仅为对照组的43%~51%(t≥3.95 7,P<0.05);增殖活性显著低于对照组(t≥3.858,P<0.0 5),且对血小板衍生生长因子(PDGF)促增殖效应的敏感性显著减弱;Ⅰ、Ⅲ型胶原和α-SMA的表达显著减少(t≥6.790,P<0.01);凋亡发生率显著高于对照组(x2≥14.157,P<0.01),电镜下可见典型凋亡细胞。 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。为抗肝纤维化治...

目的 研究抗血小板衍生生长因子受体β亚单位(PDGFR-β)核酶在肝星状细胞(HSC)内的切割活性及其对HSC生物学特性的影响。 方法 构建抗PDGFR-β核酶的真核表达载体,将其转染入HSC-T6细胞,G418筛选出阳性细胞克隆;分别用northern blot、western blot和免疫细胞化学检测PDGFR-β表达,用MTT法检测细胞增殖,免疫细胞化学检测α-F滑肌肌动蛋白(α-sMA)和Ⅰ、Ⅲ型胶原表达,用流式细胞仪、吖啶噔荧光染色和电镜分析细胞凋亡。 结果 转染核酶的HSC的PDGFR-β在mRNA和蛋白水平的表达量均显著降低,仅为对照组的43%~51%(t≥3.95 7,P<0.05);增殖活性显著低于对照组(t≥3.858,P<0.0 5),且对血小板衍生生长因子(PDGF)促增殖效应的敏感性显著减弱;Ⅰ、Ⅲ型胶原和α-SMA的表达显著减少(t≥6.790,P<0.01);凋亡发生率显著高于对照组(x2≥14.157,P<0.01),电镜下可见典型凋亡细胞。 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。为抗肝纤维化治疗提供了新的靶点和手段。

Objective To study the cleavage activity of specific deoxyribozyme to hepatitis C virus in vitro. Methods Three deoxyribozymes were designed to cleave at sites 157,168,173 in HCV 5'-noncoding region with the same active region of 5'-GGCTAGCTACAACGA-3'respectively.Plasmid pCMV/T7-NCRC-△ Luc was completely linearized with restriction endonuclease Xba I.HCV RNA5'-NCRC was transcribed in vitro from the linearized products and radiolabelled with [α-~(32) P]UTP.Under the conditions of 37℃,pH 7.5,Mg~(2+)10 mmol/L,the...

Objective To study the cleavage activity of specific deoxyribozyme to hepatitis C virus in vitro. Methods Three deoxyribozymes were designed to cleave at sites 157,168,173 in HCV 5'-noncoding region with the same active region of 5'-GGCTAGCTACAACGA-3'respectively.Plasmid pCMV/T7-NCRC-△ Luc was completely linearized with restriction endonuclease Xba I.HCV RNA5'-NCRC was transcribed in vitro from the linearized products and radiolabelled with [α-~(32) P]UTP.Under the conditions of 37℃,pH 7.5,Mg~(2+)10 mmol/L,the three deoxyribozymes were mixed with substrate RNA individually for 120 minutes and then the reactions were terminated.The cleavaged products were separated with 8% denaturated polyacrylamide gel electrophoresis and displayed by autoradiography.DRz3 was mixed with the substrate RNA at different Mg~(2+)concentrations.The cleavage efficiency was analyzed with a gel document action analyzing systems.Result Under the adopted conditions the three deoxyribozymes efficiently cleaved the target RNA in vitro and the cleavage activity of DRz3 was increased with the increase of Mg~(2+)concentration.Conclusion The designed deoxyribozymes can cleave 5'-NCR mRNA of HCV efficiently in vitro and it is dose-respondent to Mg~(2+)concentration.

目的 研究特异性脱氧核酶对丙型肝炎病毒(HCV)的体外消化作用。方法 设计3个分别 作用于 HCV RNA 5′-非编码区(5′-NCR)上157、168、173位点的脱氧核酶(DRz),分别命名为 DRzl, DRz2,DRz3,均以5′-GGCTAGCTACAAcGA-3′为脱氧核酶的催化活性中心。用内切酶 Xba Ⅰ将含有 HCV 病毒序列的质粒 pCMV/T7-NCRC-△Luc 消化成线性,以此为模板体外转录出用[α-~(32)p]UTP 进行标 记的靶 HCV RNA。在37℃、pH 7.5、Mg~(2+)10 mmol/L 等条件下,将 HCV RNA(10 nmol/L)与3个脱 氧核酶(1 μmol/L)分别混合进行切割反应,并进一步比较不同 Mg~(2+)浓度下 DRz3的切割反应效率,反应 产物在8%变性聚丙烯酰胺凝胶上电泳分离并行放射自显影,应用凝胶成像分析仪评价脱氧核酶的切割效率。 结果 在设定反应条件下,3个脱氧核酶对 HCV 病毒均有切割活性。随着 Mg~(2+)浓度的增加,DRz3的切割 活性增强。结论 特异性的脱氧核酶可有效切割 HCV RNA 5′NCR 的 RNA,且切割...

目的 研究特异性脱氧核酶对丙型肝炎病毒(HCV)的体外消化作用。方法 设计3个分别 作用于 HCV RNA 5′-非编码区(5′-NCR)上157、168、173位点的脱氧核酶(DRz),分别命名为 DRzl, DRz2,DRz3,均以5′-GGCTAGCTACAAcGA-3′为脱氧核酶的催化活性中心。用内切酶 Xba Ⅰ将含有 HCV 病毒序列的质粒 pCMV/T7-NCRC-△Luc 消化成线性,以此为模板体外转录出用[α-~(32)p]UTP 进行标 记的靶 HCV RNA。在37℃、pH 7.5、Mg~(2+)10 mmol/L 等条件下,将 HCV RNA(10 nmol/L)与3个脱 氧核酶(1 μmol/L)分别混合进行切割反应,并进一步比较不同 Mg~(2+)浓度下 DRz3的切割反应效率,反应 产物在8%变性聚丙烯酰胺凝胶上电泳分离并行放射自显影,应用凝胶成像分析仪评价脱氧核酶的切割效率。 结果 在设定反应条件下,3个脱氧核酶对 HCV 病毒均有切割活性。随着 Mg~(2+)浓度的增加,DRz3的切割 活性增强。结论 特异性的脱氧核酶可有效切割 HCV RNA 5′NCR 的 RNA,且切割效率与 Mg~(2+)浓度呈 正相关。

 
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