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暂态表达
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  transient expression
     The COS cells were transinfected by pC-IL-2 using DEAE-Dextran method for transient expression of IL-2 gene.
     采用DEAE-Dextran介导的方法,用IL-2真核表达载体pC-IL-2转染COS细胞进行暂态表达
短句来源
     TRANSIENT EXPRESSION IN MILK OF GOATS AFTER TRANSFERRING THE BOVINE αs1 CASEIN HBsAg GENE
     牛αs1酪蛋白/HBsAg基因在山羊乳腺中暂态表达
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     In order to verify cloned Calbindin-D28k cDNA encoding condition, we establish eukaryotic expression plasmid pCEP4-CaBP-D28k, which is made by Calbindin-D28k cDNA cloning into vector pCEP4. COS-1 cell carry out transient expression, which is transfectioned by pCEP4-CaBP-D28k. Immunity fiuorescein look over expressing product.
     为了验证克隆的鸡CaBP-D28k cDNA的编码情况,本研究通过将鸡CaBP-D28k cDNA亚克隆入真核表达载体pCEP4内,构建了真核表达构件pCEP4-CaBP-D28k,用所获得的真核表达构件pCEP4-CaBP-D28k转染COS-1细胞,进行暂态表达,用免疫荧光法检测到表达产物。
短句来源
     Optimization of Chicken Oviduct-Specific Expression Vectors for Transient Expression of Recombinant Proteins
     暂态表达重组蛋白鸡输卵管生物反应器的优化
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     Studies on Characteristic and Transient Expression of Antimicrobial Peptides in Mammary Glands
     抗菌肽乳腺组织特异性暂态表达及特性研究
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  “暂态表达”译为未确定词的双语例句
     Temporary Expression of Human Lysozyme cDNA in COS-1 Cells and Murine Mammary Gland
     人溶菌酶cDNA在COS-1细胞及小鼠乳腺的暂态表达
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     Transient Expression of Human Growth Hormone in Mice Milk after in vivo Transfer of hGH Gcne
     活体注射导入的人生长激素融合基因在小鼠乳腺中的暂态表达
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     Cloning,Sequencing of the Rabies Virus Nucleoprotein Structural Gene and Its Transient Expression in Cos 7 Cells
     狂犬病病毒3aG株核蛋白基因cDNA的克隆、序列分析及在COS细胞中的暂态表达
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     The possibility of producing vaccine was probed through plant virus-based gene vector expression system.
     同时本研究还通过构建LTB—ST融合基因PVX病毒表达载体,并用农感染法转化植物,探索了暂态表达系统生产疫苗的可能性。
短句来源
     These gene constructs were injected into mammary gland tissue of ewe respectively about 10 days before secreting milk and were expressed transiently.
     此六个基因构件分别在泌乳之前10天左右导入母山羊的乳腺组织中进行暂态表达
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     This expression pattern was further conformed by RT-PCR.
     到其表达
短句来源
     Abnormal expression of TGF- ?
     的表达
短句来源
     Transient Expression of Human Lysozyme in Laying Hen Oviduct
     鸡输卵管暂态表达人溶菌酶的研究
短句来源
     Generation of Chicken Oviduct Transient Bioreactor Expressing Human Lysozyme
     鸡输卵管暂态生物反应器表达人溶菌酶的研究
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     TRANSIENT MAGNETISM
     暂态磁性
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  transient expression
Construction of a bidirectional promoter and its transient expression in Populus tomentosa
      
The transient expression of the gusA and gfp genes were detected by histochemical staining for GUS and by fluorescence microscopy for GFP.
      
The direction of transient expression of GUS and GFP in Agrobacterium mediated 3-4 days transformed leaf discs of Populus tomentosa, indicating that the promoter did have bidirectional transcriptional activities simultaneously in cells and tissues.
      
The highest transient expression was observed after explant preincubation for 12-14 days and bombardment with 1 μm gold particles at the helium pressure of 61.24-74.85 atm, vacuum pressure of 0.064 atm, and distance to target of 9 cm.
      
Agroinfiltration-mediated transient expression assays showed that CRP neither acts as an avirulence factor in N.
      
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Erythropoietin expressing plasmid type adenovirus vector psp1B/hEPO was constructed.A completed EPO expression cassette promoted by RSV LTR was inserted into the Eco R Ⅴ site of the adenovirus vector pΔE1SP1B.The transient EPO expressing capability was detected by transfecting the plasmid of psp1B/hEPO into the CHO cell.By co transfecting psp1B/hEPO with the rescue type adenovirus vector—pBHG11,the recombinant adenovirus AdhEPO was obtained.EPO expressing cassette was found in AdhEPO by Southern blot...

Erythropoietin expressing plasmid type adenovirus vector psp1B/hEPO was constructed.A completed EPO expression cassette promoted by RSV LTR was inserted into the Eco R Ⅴ site of the adenovirus vector pΔE1SP1B.The transient EPO expressing capability was detected by transfecting the plasmid of psp1B/hEPO into the CHO cell.By co transfecting psp1B/hEPO with the rescue type adenovirus vector—pBHG11,the recombinant adenovirus AdhEPO was obtained.EPO expressing cassette was found in AdhEPO by Southern blot and positive EPO expression was detected using ELISA method.5×10 8 pfu of AdhEPO was administrated to the SD rat by a single intramuscular injection to investigate the EPO expression and its effect of erythropoiesis.EPO expression and polycythemia were found in a period of 10 days.The hematocrites,hematogloblin and the number of RBC(Red blood cell)of rats were detected after injections of 1,3,5,7 and 10 days.The significant increase of RBC was found.The hematocrites increased from 46±4% to 65±6% after 10 days of injection.It was found that the potential value of clinical application of recombinant adenovirus—AdhEPO could be used for gene therapy of anemia.

用EPO基因组基因构建了腺病毒质粒型载体psp1B/hEPO,该质粒含有以RSV-LTR为启动子的完整的EPO基因表达盒.单独转染CHO细胞,经暂态表达检测到EPO的表达。用psp1B/hEPO与腺病毒拯救型载体pBHG11共转染293细胞,获得了表达EPO的重组腺病毒AdhEPO.经Southern杂交证实AdhEPO中有EPO表达盒,ELISA检测到了EPO阳性表达.用5×108pfu的AdhEPO给大鼠作一次性肌肉注射,观察到了其促进大鼠红细胞生成的短期效应。在注射后第1,3,5,7,10d分别检测了大鼠的红细胞压积、血红蛋白含量和红细胞计数等指标,发现大鼠的红细胞数量显著提高。在第10d红细胞压积从46±4%上升至65±6%。证实了重组腺病毒AdhEPO具有潜在的临床应用价值,可用于贫血症的基因治疗。

Objective To let furin convert human proinsulin into mature insulin in mammalian cell.Methods Human insulin genomic gene was mutated by oligonucleotide directed mutagenesis method. Two furin recognized sites of Arg X Lys Arg motifs were introduced into human proinsulin. Expression of the mutated gene in CHO cell were carried out.Results First, A fragment of human insulin gene was cloned and connected with M13mp18 Vector so that M13mp18/INS phage vector was constructed. According to Kunkel's method,...

Objective To let furin convert human proinsulin into mature insulin in mammalian cell.Methods Human insulin genomic gene was mutated by oligonucleotide directed mutagenesis method. Two furin recognized sites of Arg X Lys Arg motifs were introduced into human proinsulin. Expression of the mutated gene in CHO cell were carried out.Results First, A fragment of human insulin gene was cloned and connected with M13mp18 Vector so that M13mp18/INS phage vector was constructed. According to Kunkel's method, by use of single strain DNA template of M13mp18/INS and two mutating primers (Ⅰ:CTGCAGGTCCTCGCGCTTCCGGCGGGTC,Ⅱ:CACGCTTCTGCCGGGATCCCTC) two sites of the gene were mutated simultaneouly. After screening and sequencing of many mutated phage vectors, a correctly mutated phage M13mp18/MINS was obtained. Second, by use of CMV promoter and the mutated insulin gene, a eukaryotic cell expression vector CMV/MINS was constructed. Transient expression of the mutated gene with CMV/MINS was carried out in CHO cell. While measuring human insulin in the medium of the expressing CHO cells by RIA method, the results of transient expression were 5 5~70 0μIU/5×10 6/cells/Day. Isolation of insulin expressing CHO cell lines were also carried out with CMV/MINS vector by G418 selection. The expression levels of the CHO strains were 6 5~25 5μIU/5×10 6 cells/Day.Conclusion A correctly mutated phage M13mp18/mINS was obtained and transfected into CHO cells with insulin expression.

目的 研究在哺乳动物细胞中表达人胰岛素。方法 采用寡核苷酸介导的定点突变方法,改造人胰岛素基因组基因,将哺乳动物细胞内蛋白质前体加工酶Furin 的识别和切割位点Arg-X-Lys-Arg-样序列,引入人胰岛素原C肽两端,在普通CHO细胞表达了该改造后的基因。结果 首先应用突变引物Ⅰ:CTGCAGGTCCTCGCGCTTCCGGCGGGTC,引物Ⅱ:CACGCTTCTGCCGGGATCCCTC,按照Kunkel的方法,成功地进行了人胰岛素基因上两个位点的同时突变改造。利用表达载体PRC/CMV和突变后的胰岛素基因,构建成表达载体CMV/MINS,并在CHO细胞中获得了表达。结论 用RIA法检测在暂态表达的CHO细胞培养液中人胰岛素的表达量为5.5~70.0μIU/5×106 细胞·d- 1。同时,还利用G418进行了胰岛素表达细胞株的筛选,筛选出的CHO细胞株的表达量为:6.5~25.5μIU/5×106细胞·d- 1。进一步需研究Furin酶对表达的人胰岛素的加工效果和表达产物的生物学活性

To pursue insulin and islet transplantation replacement therapy for type 1 diab etes based on engineered human non β cells which secrete mature insulin Methods Human proinsulin cDNA was cloned from its genomic gene and mutated by overlap e xtension PCR, introducing furin consensus cleavage sequences (Arg Xaa Lys/Arg Arg) An expression vector encoding a genetically modified human proinsulin c DNA was generated and transduced to Hela, 293, and L02 cells by lipofectin medi ated DNA transfection Following...

To pursue insulin and islet transplantation replacement therapy for type 1 diab etes based on engineered human non β cells which secrete mature insulin Methods Human proinsulin cDNA was cloned from its genomic gene and mutated by overlap e xtension PCR, introducing furin consensus cleavage sequences (Arg Xaa Lys/Arg Arg) An expression vector encoding a genetically modified human proinsulin c DNA was generated and transduced to Hela, 293, and L02 cells by lipofectin medi ated DNA transfection Following G418 screening, the surviving L02 cells were s elected and enriched Insulin levels in the supernatant and cells were evaluate d using radioimmunoassay and immunofluorescence staining Results Three sites in the insulin gene were mutated simultaneously Insulin gene m odified cells were able to express insulin at different levels: 8 45-188 00? μIU/24 h/2 0×10 6 Hela cells and 159 88-242 14?μIU/24 h/2 0×10 6 293 cells for transient expression, and 2 56-61 95?μIU/24 h/2 0×10 6 from se veral L02 clones screened with G418 No insulin was released by control cells Furthermore, immunofluorescence staining confirmed that proinsulin was stored a s vacuoles in the cytoplasm of L02 cells Conclusion A correctly mutated human proinsulin cDNA was obtained successfully, transfected and expressed efficiently in non beta cells, lending support to the study of s omatic gene therapy in diabetes mellitus

目的 研究在非β细胞中表达成熟胰岛素 ,探索替代胰岛素注射或胰岛移植治疗 1型糖尿病的方法。方法 采用重叠区扩增基因拼接法 (genesplicingbyoverlapextention ,geneSOEing)自胰岛素原基因组基因中扩增胰岛素原的cDNA ,并于C肽的两端引入蛋白酶furin的识别和切割位点Arg Xaa Lys/Arg Arg。将获得的突变体重组表达载体PcDNA3 1/C ,通过脂质体法转染体外培养的Hela ,2 93,和L0 2细胞 ,G4 18筛选L0 2细胞。同时用放射免疫和免疫荧光的方法监测细胞内外胰岛素的表达水平。结果 成功地对胰岛素原cDNA的三个位点进行了突变 ,空载体转染的细胞无胰岛素表达。而在转染突变后胰岛素原基因cDNA的细胞培养液中胰岛素的表达量各不相同 :Hela和 2 93细胞的暂态表达量分别为 8 4 5~ 188 0 0μIU/ 2 0× 10 6 cells·d 1和 15 9 88~ 2 4 2 14 μIU/ 2 0× 10 6 cells·d 1,筛选出的L0 2各细胞株的表达量为 2 5 6~ 6 1 95μIU/ 2 0×...

目的 研究在非β细胞中表达成熟胰岛素 ,探索替代胰岛素注射或胰岛移植治疗 1型糖尿病的方法。方法 采用重叠区扩增基因拼接法 (genesplicingbyoverlapextention ,geneSOEing)自胰岛素原基因组基因中扩增胰岛素原的cDNA ,并于C肽的两端引入蛋白酶furin的识别和切割位点Arg Xaa Lys/Arg Arg。将获得的突变体重组表达载体PcDNA3 1/C ,通过脂质体法转染体外培养的Hela ,2 93,和L0 2细胞 ,G4 18筛选L0 2细胞。同时用放射免疫和免疫荧光的方法监测细胞内外胰岛素的表达水平。结果 成功地对胰岛素原cDNA的三个位点进行了突变 ,空载体转染的细胞无胰岛素表达。而在转染突变后胰岛素原基因cDNA的细胞培养液中胰岛素的表达量各不相同 :Hela和 2 93细胞的暂态表达量分别为 8 4 5~ 188 0 0μIU/ 2 0× 10 6 cells·d 1和 15 9 88~ 2 4 2 14 μIU/ 2 0× 10 6 cells·d 1,筛选出的L0 2各细胞株的表达量为 2 5 6~ 6 1 95μIU/ 2 0× 10 6 cells·d 1;免疫荧光显示L0 2细胞浆内有囊泡状分泌颗粒。结论 突变的胰岛素原cDNA能成功转染非胰岛 β细胞并表达胰岛素 ,为进一步开展糖尿病的基因治疗研究奠定了基础。

 
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