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ligand-binding domains
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     The first part of the thesis describes the expression and characterization of the human insulin receptor's ligand-binding domains L1 and L2. The absence of the detailed three-dimensional structure of human insulin receptor's ectodomain remains a hindrance to understanding the molecular mechanism of insulin binding and transmembrane signaling.
     第一部分阐述了人胰岛素受体配基结合结构域L1和L2的可溶性表达并鉴定了它们的配基结合能力。 由于缺少胰岛素受体胞外部分和受体-配基复合物的详细三维结构,因此对胰岛素的结合和跨膜信号传递的分子机制仍然处于推测阶段。
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  相似匹配句对
     Osteoprotegerin and its ligand
     骨保护素及其配体
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     Ligand Behavior and the Stability of Tetrahedrane
     配体行为与四面体碳烷的稳定性
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     Ligand-independent receptor activation
     非配体依赖性受体激活
     Progress of a Proliferation-Inducing Ligand
     增殖诱导配体(APRIL)研究进展
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     Tetramethylpyrazine inhibition on binding of radiolabeled ligand to VEGFR
     川芎嗪对VEGF受体与其放射性配体结合的抑制
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  ligand-binding domains
Structure Function Studies: Identification of Vitamin D Analogs for the Ligand-Binding Domains of Important Proteins in the Vita
      
The structure of several ligand-binding domains in different ligation states has provided evidence for a ligand-dependent transcriptional switch and a molecular basis for the mode of action of agonists and antagonists.
      
Two motifs have been identified as ligand-binding domains a charged collagen structure of type I and II receptors, and an immunodominant domain of CD36.
      
Characterization of Group A Streptococcal M23 Protein and Comparison of the M3 and M23 Protein's Ligand-Binding Domains
      
Ligand-Binding Domains in Vitellogenin Receptors and Other LDL-Receptor Family Members Share a Common Ancestral Ordering of Cyst
      
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Total RNAs were extracted from human fresh placenta, and the cDNA fragment of the M CSFR extracellular region, which encoded the ligand binding domains, was amplified by RT PCR. The cDNA was then cloned into the expression vector downstream to the His Tag site. After transformation into E.coli BL21 (DE3) host bacteria, the recombinant rhM CSFsR protein was expressed efficiently in inclusion bodies with the yield of 38% total bacteria proteins. The recombinant protein was purified by affinity chromatogaphy...

Total RNAs were extracted from human fresh placenta, and the cDNA fragment of the M CSFR extracellular region, which encoded the ligand binding domains, was amplified by RT PCR. The cDNA was then cloned into the expression vector downstream to the His Tag site. After transformation into E.coli BL21 (DE3) host bacteria, the recombinant rhM CSFsR protein was expressed efficiently in inclusion bodies with the yield of 38% total bacteria proteins. The recombinant protein was purified by affinity chromatogaphy using a His Bind Resin column charged with nickel. The rhM CSFsR showed a single band in SDS PAGE and the apparent molecular weight was about 34 kD. Enzyme linked immunosorbent assay showed that rhM CSFsR had obviously specific binding activity for its ligand, the K d for dissociation was 7 8 nmol/L, and rhM CSFsR had one binding site. It suggests that glycosylation may be not essential for M CSFR. This throws a light on the studies of biological function and potential clinical application of rhM CSFsR .

从人胎盘中提取总RNA,利用RT-PCR技术扩增出巨噬细胞集落刺激因子受体(M-CSFR)胞外具有结合活性区域的cDNA,经平端连接将其克隆到原核表达载体pET-28a的His-Tag下游,转化大肠杆菌BL21(DE3)后,经IPTG诱导,重组的人可溶型M-CSFR(rhM-CSFsR)在宿主菌中获得高效表达,表达量约占菌体总蛋白的38%.重组蛋白经Ni2+组氨酸结合树脂螯合层析柱纯化,获得了纯化的rhM-CSFsR,经SDS-PAGE显示为单一区带,其表观分子量为34kD.用酶联免疫吸附分析(enzyme-linkedimmunosorbentassay,ELISA)证明rhM-CSFsR有明显的M-CSF专一结合活性,Kd值为7.8nmol/L,只有一个M-CSF结合位点.本实验结果显示原核表达的rhM-CSFsR具有明显的配基结合活性,提示M-CSFR的糖基化程度对于其配基结合活性不是必不可少的,为深入进行rhM-CSFsR的生物学功能及其临床意义的研究打下了良好的基础

AIM: All these efforts provide the basis for designing new and selective retinoid drugs. METHODS AND RESULTS: The sequence alignment based upon RAR(retinoic acid nuclear receptors ) crystal structural motifs was performed, and showed that only three amino acid residues of RAR, RAR, RAR in the ligandbinding pocket are at variance. These divergent residues are obvious features to account for the RAR selectivity of certain retinoid. Meanwhile, the ligand binding domains of holoRAR(,) were modeled by site...

AIM: All these efforts provide the basis for designing new and selective retinoid drugs. METHODS AND RESULTS: The sequence alignment based upon RAR(retinoic acid nuclear receptors ) crystal structural motifs was performed, and showed that only three amino acid residues of RAR, RAR, RAR in the ligandbinding pocket are at variance. These divergent residues are obvious features to account for the RAR selectivity of certain retinoid. Meanwhile, the ligand binding domains of holoRAR(,) were modeled by site mutagenesis technique. Molecular dynamics relaxing and global minimization were carried out and led to construction of the final complex models. The interaction of RAR(,,) and their specific ligand by docking simulation were investigated, the fine binding patterns have been used to define clearly some important structural characteristics of selective retinoids. CONCLUSION: Receptorspecific retinoids possess improved activity/toxicity profiles compared with the nonselective counterparts. Molecul

目的:研究维甲酸核受体蛋白α,β和γ3种亚型的氨基酸序列差异性,确定可能与抑制剂选择性结合有关的氨基酸残基,揭示维甲酸类药物的毒副作用和与其结合多种受体亚型的关系,设计、开发受体选择性的维甲类分子。方法和结果:基于已解析的γ受体亚型的晶体结构,利用分子图形学和计算化学方法预测了未知的α,β亚型的三维结构;通过3种维甲酸核受体分别与其泛激动剂、选择性激动剂及无结合活性的维甲酸分子之间的分子对接研究,定性解析了抑制剂立体结构上的微小差异导致不同受体产生选择性的本质原因。结论:有选择性受体结合活性的维甲类化合物是进行低毒作用的新型维甲类药物研究和开发的基础。

ObjectiveTo screen and identify the bioactive peptides that bind to tyrosine kinase receptor EphB2.MethodsThe gene encoding the ligand binding domain of EphB2 was cloned into the expressing vector pRSET A,under induction with IPTG,the fusion protein was expressed and purified under denaturing condition using metal chelate chromatography.The purified protein coated on ELISA plate was used as target to carry out three rounds of affinity selection.ResultsSDS...

ObjectiveTo screen and identify the bioactive peptides that bind to tyrosine kinase receptor EphB2.MethodsThe gene encoding the ligand binding domain of EphB2 was cloned into the expressing vector pRSET A,under induction with IPTG,the fusion protein was expressed and purified under denaturing condition using metal chelate chromatography.The purified protein coated on ELISA plate was used as target to carry out three rounds of affinity selection.ResultsSDS PAGE analysis showed that the fusion protein mainly existed in inclusion bodies.The purity was up to 95%,13 positive phages were isolated from a random phage displayed twelve peptide library.ConclusionPeptides that could interact specifically with EphB2 were obtained and there were common motifs among the sequences of them.

目的筛选能结合酪氨酸蛋白激酶受体 Eph B2的功能短肽。方法 PCR扩增 Eph B2的配体结合区 ,定向克隆到融合表达质粒载体 p RSET A中 ,阳性克隆经 IPTG诱导表达融合蛋白 ,并利用 Ni- NTA金属螯合亲和层析在变性条件下对表达的蛋白进行纯化 ,以此纯化蛋白为靶 ,将其包被于 EL ISA板上 ,进行 3轮亲和筛选。结果电泳分析表明 :表达的融合蛋白主要以包涵体的形式存在 ,纯化蛋白质的纯度大于 95 %。从噬菌体随机 12肽库中筛选到 13个噬菌体阳性克隆。结论获得了具有结合 Eph B2活性的阳性噬菌体克隆且有共同的基序

 
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