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functional dna
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     THE CLONING OF A FUNCTIONAL DNA FRAGMENT OFRA538 GENE AND ITS EFFECT ON HL-60 CELLS
     RA538基因功能片段的克隆和对HL-60细胞的作用
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     Although the biological significance of DNA methylation has been recognized to some extent, the mechanisms of DNA methylation regulation and of the establishment of tissue-specific DNA methylation pattern in somatic cells are still unclear, since the knowledge of the enzymes responsible for de novo DNA methylation and demethylation of DNA is very limited and up to now only two functional DNA methyltransferases in mammals have been cloned .
     目前,虽然对DNA甲基化的作用已有一定认识,但DNA甲基化的调节以及发育过程中组织特异的DNA甲基化谱的建立机制还不清楚。 问题主要在于对催化重新甲基化和去甲基化反应的酶的了解很少。
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     Methods The cultured cells were transfected with the functional DNA cassettes that contained RNA polymers Ⅲ promotor,small hairpin interference RNA template and transcription termination sequence,then were stimulated by lipopolysaccharide(LPS).
     方法体外合成针对TNFα基因序列的特异性发夹状双链RNA(shRNA)的表达载体,与L ipofectam ine2000结合后转染原代培养的由脂多糖(LPS)激活的脾淋巴细胞,用ELISA和RT-PCR法检测TNFα表达的变化。
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     DNA Polymorphism and Its Applications in Plant Functional Genomics
     DNA多态性及其在植物功能基因组学研究中的应用
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     Study on the Functional Domain of Taq DNA Polymerase
     Taq DNA聚合酶功能区域的定位
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     On Classification of DNA
     DNA分类方法的探讨
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     DNA nanotechnology
     DNA纳米技术
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     Functional Egg
     功能性鸡蛋
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  functional dna
Together, these data indicate that we have cloned the canine AR and that its functional DNA binding and ligand binding domains are absolutely conserved with those reported for eight other species.
      
Southern blot analysis of inserted genes in transgenic rice plants suggests the integration of an intact hygromycin phosphotransferase gene and non-functional DNA fragments into host genome.
      
Interestingly, each ZmHox2 gene product contains two complete homeodomains which, for Zmhox2a, were both shown to be functional DNA-binding motifs in vitro.
      
These findings indicatethat the longevity hormesis may be associatedwith irradiation-induced long-term structuraland/or functional DNA modifications.
      
This review examines evolution of bacterial genomes with an emphasis on RNA based life, the transition to functional DNA and small evolving genomes (possibly plasmids) that led to larger, functional bacterial genomes.
      
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Seven promoter-functional DNA fragments of different sizes have been cloned from Penicillium decumbens JUA-10 by using promoter-detecting plasmid pSUPV4 as vector and E.coli JM109 as host strain.The result of the experiment on kanamysin resistance indicaties that all of the seven E.coli Kan.transformants are able to grow in the medium containing kanamysin of 100 μg/ml~1100μg/ml,which demonstrates that some promoter-functional DNA fragments from Penicillium decumbens are active in...

Seven promoter-functional DNA fragments of different sizes have been cloned from Penicillium decumbens JUA-10 by using promoter-detecting plasmid pSUPV4 as vector and E.coli JM109 as host strain.The result of the experiment on kanamysin resistance indicaties that all of the seven E.coli Kan.transformants are able to grow in the medium containing kanamysin of 100 μg/ml~1100μg/ml,which demonstrates that some promoter-functional DNA fragments from Penicillium decumbens are active in E.coli .A further Restriction enzyme analysis of plasmid pPdsu1 with the highest level of resistance to kanamysin is conducted and the result indicated that the length of the fragment inserted into pPdsu1 is of 2.4kb.Result of Southem hybridization analysis indicates that this DNA fragment originally comes from Penicillium decumbens chromosomal DNA.

 将斜卧青霉JUA - 10染色体DNA的BamHI或PstI片段连接到大肠杆菌的启动子探针型载体pSUPV4上 ,克隆了 7个不同大小的具有启动子功能的DNA片段。卡那霉素抗性试验表明 ,含重组质粒的大肠杆菌均能耐受 10 0 μg/ml以上的卡那霉素 ,最高可达 110 0 μg/ml。说明斜卧青霉的某些DNA片段能在大肠杆菌中有效地启动基因表达。取抗性水平最高的 pPdsuI进一步进行限制酶谱分析 ,表明插入片段的分子量为 2 .4Kb。Southern杂交分析结果表明该插入DNA片段与斜卧青霉染色体DNA具同源性

Objective To investigate the effect of small hairpin interference RNA(shRNA)on the expression of tumor necrosis factoralpha(TNFα) in primary cultured spleen lymphocytes.Methods The cultured cells were transfected with the functional DNA cassettes that contained RNA polymers Ⅲ promotor,small hairpin interference RNA template and transcription termination sequence,then were stimulated by lipopolysaccharide(LPS).TNFα expression was assessed by ELISA and RT-PCR.Results Sequence specific siRNA targeting TNFα...

Objective To investigate the effect of small hairpin interference RNA(shRNA)on the expression of tumor necrosis factoralpha(TNFα) in primary cultured spleen lymphocytes.Methods The cultured cells were transfected with the functional DNA cassettes that contained RNA polymers Ⅲ promotor,small hairpin interference RNA template and transcription termination sequence,then were stimulated by lipopolysaccharide(LPS).TNFα expression was assessed by ELISA and RT-PCR.Results Sequence specific siRNA targeting TNFα could down-regulate the expression of TNFα significantly.Compared with the control group,gene specific shRNA-TNFα reduced TNFα expression by 34.2%,and the nonspecific interference sequence had no influence on TNFα expression,gene specific shRNA-TNFα inhibited TNFα mRNA expression by 61.3%.Conclusion Sequence specific shRNA targeting TNFα could effectively inhibit the expression of TNFα in the primary cultured lymphocytes.This successful application extends the list of available therapeutic modalities in the treatment of diseases involed TNFα.

目的观察RNA干扰技术是否有效抑制原代培养的脾淋巴细胞肿瘤坏死因子α(TNFα)的表达。方法体外合成针对TNFα基因序列的特异性发夹状双链RNA(shRNA)的表达载体,与L ipofectam ine2000结合后转染原代培养的由脂多糖(LPS)激活的脾淋巴细胞,用ELISA和RT-PCR法检测TNFα表达的变化。结果ELISA检测结果表明,特异性RNA干扰组TNFα蛋白表达较对照组下降34.2%,而非特异性干扰组TNFα表达与对照组无明显差异;RT-PCR检测结果表明,特异性RNA干扰组TNFαmRNA表达较对照组下降61.3%。结论RNA干扰可以有效抑制原代培养的淋巴细胞TNFα的表达,该过程具有严格的基因序列特异性,为TNFα相关疾病的基因治疗提供了新策略。

 
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