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binding plasmid
相关语句
  质粒结合
     Purpose To express and purify the fused protein containing 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines, and investigate its ability of binding plasmid.
     目的 表达和纯化流感病毒血凝素HA2氨基末端20个氨基酸与10个赖氨酸的融合蛋白HA20-lys10,并观察其与质粒结合的能力。
短句来源
  “binding plasmid”译为未确定词的双语例句
     (4)Deposition experiment showed that the directional movement of pll-DCIONP binding plasmid under magnetic field in liquid.
     (4)沉淀实验证明,pll-DCIONP 可携带质粒DNA 在外加磁场作用下在液体中定向移动。
短句来源
     The expressed protein was purified by one-step affinity chromatography, and its ability of binding plasmid was investigated via gel retardation experiments.
     表达产物用亲和层析一步法进行纯化,然后用凝胶阻滞试验观察重组蛋白结合质粒的能力。
短句来源
     Deposition experiment shows that the directional movement of pll DCIONP binding plasmid under magnetic field in liquid.
     沉淀实验证明pll DCIONP可携带质粒DNA在外加磁场作用下在液体中定向移动。
短句来源
  相似匹配句对
     the plasmid was transformed into E.
     coli JM 109扩增此质粒后经CpG甲基化酶M.
短句来源
     The recombinant plasmid of E.
     应用细菌质粒转化技术,将大肠杆菌K_(?)
短句来源
     protein binding;
     蛋白结合;
短句来源
     Binding capacity and stability of antibody and plasmid DNA on stents were quantified by radioactive labeling.
     用125I标记抗DNA抗体,检测支架表面偶联的抗体量。
短句来源
     The binding stability of plasmid was significantly better than the control groups(P<0.01).
     实验组胶原基质携带质粒DNA量显著高于对照组(P<0.01),释放时间达13d以上。
短句来源
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  binding plasmid
A 130-kDa outer membrane protein was synthesized by the transformant (pET4) carrying a 4-kb Congo red binding plasmid DNA fragment of pCAT120.
      


The fused gene ( Lys 10 PAI 1)of ten lysine and plasminogen activator inhibitor 1 was cloned into prokaryotic expression vector pET32a(+) and pET28a(+). The recombinant expression plasmid pET32 PAI and pET28 PAI were transformed into E.coli BL21(DE3). After induction for 3 hours by IPTG, the expression recombinants showed a high level expression of recombinant protein with both Trx·PAI 1 and rPAI 1 being more than 20% of the total bacterial protein. SDS PAGE and Western...

The fused gene ( Lys 10 PAI 1)of ten lysine and plasminogen activator inhibitor 1 was cloned into prokaryotic expression vector pET32a(+) and pET28a(+). The recombinant expression plasmid pET32 PAI and pET28 PAI were transformed into E.coli BL21(DE3). After induction for 3 hours by IPTG, the expression recombinants showed a high level expression of recombinant protein with both Trx·PAI 1 and rPAI 1 being more than 20% of the total bacterial protein. SDS PAGE and Western blot analysis indicated that the molecular weight of Trx·PAI 1 and rPAI 1 was about 63 kD and 43 kD, respectively and the recombinant proteins could react specifically with anti PAI 1 antibody. The purified recombinant Trx·PAI 1 and rPAI 1 were obtained after denaturation, renaturation, dialysis and affinity chromatography. In gel retardation experiment, the purified protein retarded plasmid in gel electrophoresis, and obviously changed the mobility of plasmid. These results indicate that Trx·PAI 1 and rPAI 1 can bind plasmid, and are hopeful to be used to transfer gene into cells.

用PCR方法构建 10个赖氨酸与纤溶酶原激活剂抑制物 1的融合基因Lys10 PAI 1,并克隆于pET2 8a(+ )和pET32a(+ )原核表达载体 .将重组表达质粒pET32 PAI和pET2 8 PAI转化大肠杆菌BL2 1(DE3) .IPTG诱导后可获得分子量分别为 6 3kD和 4 3kD的目的蛋白 ,表达蛋白占菌体蛋白2 0 %以上 ,大多数重组蛋白以不溶形式存在 .表达产物经变性、复性、超滤、透析和亲和层析等步骤 ,可以得到纯化的Trx·PAI 1和rPAI 1重组蛋白 .Western印迹结果表明 ,目的蛋白具有人PAI 1的抗原性 .凝胶阻滞实验发现 ,纯化的重组蛋白在一定条件下 ,可以与质粒结合 ,使质粒的迁移率明显改变 .研究结果表明 ,Trx·PAI 1和rPAI 1有希望成为受体介导基因转移的配体

Purpose To express and purify the fused protein containing 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines, and investigate its ability of binding plasmid. Methods The fused gene HA20-lys10 containing gene of 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines was amplified by PCR, and was cloned into prokaryotic expression vector pET32a( + ) . The re-combinant expression plasmid pET-HA-K was transformed into E. coli BL21 (DE3). When A600 reached...

Purpose To express and purify the fused protein containing 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines, and investigate its ability of binding plasmid. Methods The fused gene HA20-lys10 containing gene of 20 residues of amino-terminal of influenza hemagglutinin HA2 and ten lysines was amplified by PCR, and was cloned into prokaryotic expression vector pET32a( + ) . The re-combinant expression plasmid pET-HA-K was transformed into E. coli BL21 (DE3). When A600 reached 0. 6, IPTG was added to induce the expression of recombinant protein. The expressed protein was purified by one-step affinity chromatography, and its ability of binding plasmid was investigated via gel retardation experiments. Results After induction for 3 hours by 1 mmol/L IPTG, there was a high level expression of recombinant protein, and most of them were soluble. After purification by affinity chromatography, the purity of HA20-lys10 was up to 85 % , and 20μg HA20-lys10 could retard plasmid completely. Conclusions These results indicate that HA20-lys10 is hopeful to be used in receptor-mediated gene transfer to improve the efficiency.

目的 表达和纯化流感病毒血凝素HA2氨基末端20个氨基酸与10个赖氨酸的融合蛋白HA20-lys10,并观察其与质粒结合的能力。方法 用PCR方法构建流感病毒血凝素HA2氨基末端20个氨基酸与10个赖氨酸的融合基因HA20—lys10,并克隆于pET32a(+)原核表达载体。将重组表达质粒pET-HA-K转化大肠埃希菌BL21(DE3),当A(Abs)_(600nm)达到0.6时,加入终浓度为1mmol/L IPTG诱导目的蛋白的表达。表达产物用亲和层析一步法进行纯化,然后用凝胶阻滞试验观察重组蛋白结合质粒的能力。结果 IPTG诱导后可获得相对分子质量约为27 000的目的蛋白,表达蛋白以可溶形式存在,占菌体蛋白20%以上。表达菌超声后,表达产物经亲和层析一步纯化后可获得纯度达85%以上目的蛋白。凝胶阻滞试验发现,纯化的重组蛋白在一定条件下可以与质粒结合,使质粒的迁移率明显改变。结论 融合蛋白HA20—lys10有望用于受体介导基因转移,以提高基因转移的效率。

Objective To Construct Na + H + exchanger 1 (NHE 1) antisense gene magnetic nanoparticies,and evaluate feasibility of using magnetic iron oxide nanoparticies as vector to transfect NHE 1 antisense gene in therapy of cancer. Methods The dextran coated iron oxide nanoparticles (DCIONP) was synthesized with deposition. The configuration and diameter of DCIONP was detected by transmission electron microscope and zetasizer. The potential of adsorbing NHE 1 antisense gene and resisting DNase I digestion of...

Objective To Construct Na + H + exchanger 1 (NHE 1) antisense gene magnetic nanoparticies,and evaluate feasibility of using magnetic iron oxide nanoparticies as vector to transfect NHE 1 antisense gene in therapy of cancer. Methods The dextran coated iron oxide nanoparticles (DCIONP) was synthesized with deposition. The configuration and diameter of DCIONP was detected by transmission electron microscope and zetasizer. The potential of adsorbing NHE 1 antisense gene and resisting DNase I digestion of DCIONP was analyzed by agarose gel electrophoresis. The potential of orientation movement of iron oxide nanoparticles binding NHE 1 antisense gene was analyzed by attracting of magnetic field. Results Transmission electron microscope shows: nanoparticies's shape is irregular,and there is a core of ferric oxide inside nanoparticies. Under zetasizer, the mean diameter of the DCIONP is 47 nm. Agarose gel electrophoresis of binding experiment shows that pll DCIONP have the satisfactory potential to adsorb DNA.Protection experiment shows that pll DCIONP can protect DNA from DNaseI digestion effectively. Deposition experiment shows that the directional movement of pll DCIONP binding plasmid under magnetic field in liquid. Conclusions NHE 1 antisense gene magnetic nanoparticies have be constructed successfully. Small diameter, fine stability and magnetic target movement are characteristics of NHE 1 antisense gene magnetic nanoparticies.

目的 构建NHE 1反义基因磁性纳米颗粒 ,探讨以氧化铁磁性纳米颗粒为载体转染NHE 1反义基因治疗肿瘤的可能性。方法 应用共沉淀法制备外包葡聚糖的氧化铁磁性纳米颗粒(dextrancoatedironoxidenanoparticles ,DCIONP)。使用透射电镜和激光粒度检测仪检测DCIONP的形态和粒径 ,采用琼脂糖凝胶电泳分析氧化铁磁性纳米颗粒结合NHE 1反义基因及抵抗DNaseⅠ消化的能力 ,采用磁场吸引的方法分析氧化铁磁性纳米颗粒携带NHE 1反义基因定向运动的能力。结果 透射电镜观察显示 :纳米颗粒形状不规则 ,颗粒内部有一氧化铁的核心。激光粒度检测仪检测证实DCIONP的平均直径在 4 7nm左右。结合实验的琼脂糖凝胶电泳显示 ,pll DCIONP在各种质量比均有良好的DNA结合能力。DNA保护实验显示从质量比 0 .5∶1开始pll DCIONP能有效保护DNA不被DNaseⅠ降解。沉淀实验证明pll DCIONP可携带质粒DNA在外加磁场作用下在液体中定向移动。结论 成功构建了NHE 1反义基因磁性纳米颗粒 ,该颗粒具有粒径小、稳定性高及磁靶向运动的特点。

 
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