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decorin gene
相关语句
  核心蛋白聚糖基因
     Influences of Recombinant Adeno-associated Virus-mediated Decorin Gene Transfection on Cell Cycle and Apoptosis of SiHa Cells
     重组腺相关病毒介导核心蛋白聚糖基因转染对SiHa细胞周期和凋亡的影响
短句来源
  “decorin gene”译为未确定词的双语例句
     Cloning and expressing of Decorin gene
     饰胶蛋白(Decorin)基因的cDNA克隆与表达
短句来源
     Influence on the expression of type I and type II transforming growth factor-β receptors in cultured rat mesangial cells transfected by decorin gene
     饰胶蛋白聚糖基因转染对大鼠肾系膜细胞转化生长因子βⅠ、βⅡ型受体表达的影响
短句来源
     Methods The Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19.The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method,and then it was subcloned into expression plasmid vector pGEX 4T 1,the recombinant vector was indentified with BamHI and EcoRI in 1.2% agarose gel electrophoresis.
     方法 应用PCR技术从T细胞cDNA文库获得饰胶蛋白全长基因cDNA片段 ,并克隆到pUC1 9载体中 ,采用Sanger双脱氧末端终止法测定全部cDNA序列 ,将测序正确的饰胶蛋白cDNA序列插入pGEX 4T 1表达载体中 ,经BamHI和EcoRI双酶切后 1 2 %凝胶电泳鉴定 ,IPTG诱导表达产物经SDS PAGE分析。
短句来源
     The transfected tumor cells with decorin gene failed to generate tumors in scid mice, the expression of decorin mRNA and protein is various in different tumors tissues.
     国内外研究资料显示,Decorin基因及蛋白分子在不同肿瘤组织中表达是多样化的。
短句来源
     The expression of DCN is driven by 5′-LTR promoter of pLXSN. CONCLUSION An eukaryotic expression vector and retrovirus vector of decorin gene are constructed and identified that might further promote the research of mechanisms and gene therapies for tissue fibrosis.
     结论 :本研究所构建的真核表达载体pcDNA3 DCN在体外COS 7细胞可短暂表达DCN ,并成功地采用逆转录病毒载体pLXSN(克隆位点在 5′ 病毒长末端区 (5′ LTR)的下游 ,插入的目的基因受 5′ LTR启动子调控 ) ,通过EcoRI和XhoI酶切位点插入而获得核心蛋白聚糖逆转录病毒载体pLXSN DCN。
短句来源
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  相似匹配句对
     Cloning and expressing of Decorin gene
     饰胶蛋白(Decorin)基因的cDNA克隆与表达
短句来源
     CONSTRUCTION AND IDENTIFICATION OF RAT DECORIN GENE EXPRESSION VECTORS
     核心蛋白聚糖真核表达载体及逆转录载体的构建及鉴定
短句来源
     Gene Therapy
     基因治疗
短句来源
     Rescue Gene
     抢救植物基因
短句来源
     Decorin and Fibrosis
     核心蛋白聚糖与纤维化
短句来源
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  decorin gene
This review will concentrate on decorin's function in collagen fibrillogenesis as determined through the study of mice with a disrupted decorin gene.
      
This regulatory domain is likely to be implicated in the silencing of decorin gene by TGF-β and may contribute to the regulation of this matrix gene in the tumor stroma.
      
Furthermore, a transforming growth factor beta (TGF-β)-negative element is present in the promoter region of decorin gene.
      
Elucidation of decorin gene structure has revealed an unexpected degree of complexity in the 5' untranslated region of the gene with two leader exons that are alternatively spliced to the second coding exon.
      
These changes correlate with a marked increase in decorin mRNA levels and a concurrent hypomethylation of decorin gene, a DNA alteration associated with enhanced gene expression.
      
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Objective To study Decorin biological effects,Decorin gene cDNA was cloned and expressed in E.coli.Methods The Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19.The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method,and then it was subcloned into expression plasmid vector pGEX 4T 1,the recombinant vector was indentified with BamHI and EcoRI in 1.2% agarose gel electrophoresis.The expression...

Objective To study Decorin biological effects,Decorin gene cDNA was cloned and expressed in E.coli.Methods The Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19.The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method,and then it was subcloned into expression plasmid vector pGEX 4T 1,the recombinant vector was indentified with BamHI and EcoRI in 1.2% agarose gel electrophoresis.The expression of Decorin fusion protein with GST in E.coli JM109 was induced with IPTG and identified by SDS PAGE.Results The PCR amplified DNA fragment shared identical sequence with known Decorin gene reported in GenBank(Accession number:AF138300).The SDS PAGE analysis revealed that the expressed fusion protein MW was approximatelly 65.6KD and in soluble format. Conclusion The cloned Decorin gene is correct and it was expressed in fusion protein with GST.

目的 通过基因克隆技术获得饰胶蛋白基因cDNA克隆并表达。为研究饰胶蛋白的生物学功能及应用打基础。方法 应用PCR技术从T细胞cDNA文库获得饰胶蛋白全长基因cDNA片段 ,并克隆到pUC1 9载体中 ,采用Sanger双脱氧末端终止法测定全部cDNA序列 ,将测序正确的饰胶蛋白cDNA序列插入pGEX 4T 1表达载体中 ,经BamHI和EcoRI双酶切后 1 2 %凝胶电泳鉴定 ,IPTG诱导表达产物经SDS PAGE分析。结果 克隆的饰胶蛋白cDNA序列与GenBank中AF1 3830 0记载的序列完全一致 ,所表达的融合蛋白质分子量与理论计算值(65 6KD)也一致并为可溶性蛋白。结论 本研究成功地克隆了饰胶蛋白基因和表达了GST Decorin融合蛋白

Decorin(DCN)is one of the small dermatan sulfate proteoglycans and belongs to the family of leucine-rich proteoglycans of extracellular matrix.It can bind to TGF-β and thus neutralizing the fibrogenic effects of this cytokine that is implicated in the pathogenesis of progressive renal diseases.The purpose of this study is to construct eukaryotic expression vector carrying rat decorin cDNA and observe the expression of deocrin in cultured...

Decorin(DCN)is one of the small dermatan sulfate proteoglycans and belongs to the family of leucine-rich proteoglycans of extracellular matrix.It can bind to TGF-β and thus neutralizing the fibrogenic effects of this cytokine that is implicated in the pathogenesis of progressive renal diseases.The purpose of this study is to construct eukaryotic expression vector carrying rat decorin cDNA and observe the expression of deocrin in cultured COS-7 cells,and to construct retrovirus vector expressing decorin for exploring the possibility of gene therapy by decorin expression in renal diseases. METHODOLOGY Decorin cDNA with secretive signal peptide was cloned into vector pBluescript Ⅱ for sequencing,and then into eukaryotic expression vector pcDNA3 to construct pcDNA3-DCN vector,in which the transcription of decorin cDNA was driven by the CMV promoter.The vector was identified and transfected into COS-7 cells by lipofectamine.Protein secretion of decorin was detected by ELISA at 48,72 and 96 hours.Retrovirus vector pLXSN was used to construct pLXSN-DCN recombinant vector which was identified by restriction enzyme analysis(EcoR I and Xho I). RESULTS Rat DCN cDNA was inserted into pBluescript Ⅱ by enzyme cutting sites of Hind Ⅲ and Cla I.The sequence of DCN cDNA cloned in pBluescript Ⅱ-DCN met the result of Genebank.The eukaryotic expression vector pcDNA3-DCN was successfully constructed,and DCN expression was driven by CMV promoter.Then pcDNA3-DCN was successfully transfected into COS-7 cells by lipofectamin.Protein expression of decorin was increased in the supernatant of COS-7 cells transfected with pcNDA3-DCN at 72 hours after transfection as compared to the control group transfected with pcDNA3( P <0 05),pLXSN-DCN retrovirus vector was constructed by inserting enzyme cutting sites of EcoR I and Xho I and confirmed by restriction enzyme (EcoR I and Xho I)digestion.The gene of DCN is about 1 1kb.The expression of DCN is driven by 5′-LTR promoter of pLXSN. CONCLUSION An eukaryotic expression vector and retrovirus vector of decorin gene are constructed and identified that might further promote the research of mechanisms and gene therapies for tissue fibrosis.

目的 :通过获取核心蛋白聚糖 (decorin ,DCN)基因 ,构建DCN真核表达载体和DCN逆转病毒载体 ,以探索今后肾脏疾病基因治疗的途径。  方法 :从大鼠肾脏组织中抽取RNA ,经逆转录 PCR方法 ,扩增核心蛋白聚糖cDNA ,并经序列测定正确后 ,将DCN插入真核表达载体pcDNA3 ,应用Lipofectamine介导将pcDNA3 DCN转染真核细胞COS 7细胞 ,并于转染后 48、72和 96h用ELISA方法测定细胞上清液DCN的表达量 ;进一步将DCN插入逆转录病毒载体pLXSN ,抽提质粒 ,应用EcoRI和XhoI双酶切 ,选出阳性克隆 ,经大量质粒抽提、纯化后进一步鉴定插入DNA片段的正确性。  结果 :测序载体pBluescriptⅡ与DCN连接后检测DCN序列结果正确 ,从而获得DCN基因序列 ;经Lipofectamine介导转染COS 7细胞 ,于转染后 72h细胞上清液DCN的蛋白量明显增加。pLXSN DCN酶切后 ,电泳可见 1 1kb的条带 ,提示DCN插入pLXSN成功。  结论 :本研究所构建的真核表达载体pcDNA3 DCN在体外COS 7细胞可短暂表达...

目的 :通过获取核心蛋白聚糖 (decorin ,DCN)基因 ,构建DCN真核表达载体和DCN逆转病毒载体 ,以探索今后肾脏疾病基因治疗的途径。  方法 :从大鼠肾脏组织中抽取RNA ,经逆转录 PCR方法 ,扩增核心蛋白聚糖cDNA ,并经序列测定正确后 ,将DCN插入真核表达载体pcDNA3 ,应用Lipofectamine介导将pcDNA3 DCN转染真核细胞COS 7细胞 ,并于转染后 48、72和 96h用ELISA方法测定细胞上清液DCN的表达量 ;进一步将DCN插入逆转录病毒载体pLXSN ,抽提质粒 ,应用EcoRI和XhoI双酶切 ,选出阳性克隆 ,经大量质粒抽提、纯化后进一步鉴定插入DNA片段的正确性。  结果 :测序载体pBluescriptⅡ与DCN连接后检测DCN序列结果正确 ,从而获得DCN基因序列 ;经Lipofectamine介导转染COS 7细胞 ,于转染后 72h细胞上清液DCN的蛋白量明显增加。pLXSN DCN酶切后 ,电泳可见 1 1kb的条带 ,提示DCN插入pLXSN成功。  结论 :本研究所构建的真核表达载体pcDNA3 DCN在体外COS 7细胞可短暂表达DCN ,并成功地采用逆转录病毒载体pLXSN(克隆位点在 5′ 病毒长末端区 (5′ LTR)的下游 ,插入的目的基因受 5′ LTR启动子调控 ) ,通过EcoRI和XhoI酶切位点插入而获得核心蛋白聚糖逆转录病毒载体pLXSN DCN。

Purpose To study the possibility of transferring decorin gene to rat glomerular mesangial cell. Methods Amplification of the rat decorin(DCN) cDNA by RT PCR for constructing the plasmid pcDNA3.1A DCN and lipofectin method for transfecting DCN gene into MsC;G418 scanning,Western blot and RT PCR analysis for detecting DCN protein and mRNA in D A6 cell clone. Results The recombinant eukaryotic expression plasmid,pcDNA3.1A DCN was successfully constructed and 2 cell clones positively...

Purpose To study the possibility of transferring decorin gene to rat glomerular mesangial cell. Methods Amplification of the rat decorin(DCN) cDNA by RT PCR for constructing the plasmid pcDNA3.1A DCN and lipofectin method for transfecting DCN gene into MsC;G418 scanning,Western blot and RT PCR analysis for detecting DCN protein and mRNA in D A6 cell clone. Results The recombinant eukaryotic expression plasmid,pcDNA3.1A DCN was successfully constructed and 2 cell clones positively expressing DCN were selected. Conclusions These cell clones positively expressing DCN is valuable for providing favorable experimental bases of gene therapy to model with glomerular diseases.

目的 研究将饰胶蛋白聚糖 (decorin ,DCN)基因转染大鼠肾小球系膜细胞的可行性。方法 RT PCR法扩增大鼠DCN基因 ,构建重组真核表达质粒 pcDNA3.1A DCN ,采用脂质体将其转入大鼠MsC ,经G418筛选阳性克隆 ,Westernblot及RT PCR法鉴定。结果 成功构建重组真核表达质粒 pcDNA3 .1A DCN ,转染MsC并筛选出 2个阳性克隆株。结论 构建载有DCN基因的MsC载体 ,为其开展对肾小球疾病模型的基因治疗提供了良好的实验基础

 
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