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pcineo
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  pci-neo
     Mathods: pCI-AXI, the human expression vector of Annexin I was constructed by digesting the Annexin I cDNA containing plasmid pT7T3D (the American Type culture Collection) by Not I and Xho I,directional subcloning into mammalian expression vector pCI-neo containing the CMV promotor.
     方法:pT7T3D质粒经NotⅠ和XhoⅠ双酶切获得人AnnexinⅠ基因cDNA片段,亚克隆至真核表达载体pCI-neo,构建人AnnexinⅠ基因其核表达载体pCI-AXI。
短句来源
     Methods At first, pcDNA3.1 plasmid vector was digested with NheI and EcoRI to get the coding sequence of the small (S) surface Protein of HBV, then cloned into pCI-neo vector and named it pCI-S.
     方法我们首先酶切pcDNA3.1-S获得目的基因HBSAg片段,将其克隆到pCI-neo载体得到pCI-S表达载体;
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  “pci-neo”译为未确定词的双语例句
     Expression plasmid (pHCV-neo4) of luciferase controlled by HCV 5′NCR was confirmed by PCR, endonuclease digestion as well as luciferase transient expression.
     将HCV5'NCR-C基因与荧光素酶基因的融合基因片段插入pCI-neo表达载体,通过PCR扩增、酶切反应、质粒大小检测及荧光素酶瞬间表达活性鉴定等试验,获得HCV5'NCR调控荧光素酶的稳定表达质粒pHCV-neo4。
短句来源
     colt EcoR Ⅱmethylase (catalyze CCWGG→Cm CWGG) gene was recombined into eucaryotic expression vectorPCI-neo after site-directed mutagenesis, and with it mouse NIH3T3 cells were trans feeted byliposome mediation. Effectively expressing cell line was obtained through G418 selection. MostEcoR Ⅱ sites in genomic DNA of this cell line had been specifically methylated.
     来自大肠杆菌的EcoRⅡ甲基化酶(催化CCWGG→CmCWGG)基因经定点诱变后重组到真核表达载体pCI-neo上,采用脂质体介导的方法将其转染小鼠NIH3T3细胞,经G418筛选获得有效表达的细胞株,酶切证实该细胞株基因组DNA上的大部分EcoRⅡ识别位点已被甲基化,PCR鉴定EcoRⅡ甲基化酶基因已稳定整合到细胞染色体上。
短句来源
     Methods The cDNA encoding human membrane cofactor protein(hMCP) was cloned from human placenta total RNA by RT PCR . It was inserted into pCI neo mammalian expression vector and pEF neo vector which was derived from pCI neo vector. Expression plasmids pCIM and pEFM were got.
     方法从人胎盘组织提取总RNA,用RT-PCR技术扩增得到人膜辅助因子蛋白(hMCP)基因全长cDNA,将其克隆到带有巨细胞病毒(CMV)IE启动子的pCI-neo哺乳动物表达载体和以pCI-neo为基础构建成的带有人EF-1α启动子的pEF-neo哺乳动物表达载体上,得到质粒pCIM和pEFM。
短句来源
     More pCI-AXl went into S phase and Q/M phase by flow cytometric analysis, they also showed increased clonogenicity in plating efficiency (P<0.002) and soft agarose assays.
     永生化细胞、pCI-neo和pCI-AXI转染三种BEP2D细胞的接种效率分别为49.4%,57.87%,82.53%,pCI-AXI转染细胞形成的集落数明显高于其它两种细胞(P<0.002),并且细胞呈复层生长趋势;
短句来源
  相似匹配句对
     The human OSM cDNA was inserted into the EcoR I-Xba I site of the vector pCI-neo to form the OSM expression vector (pO).
     将人OSMcDNA插入 pCI-neo构成OSM表达质粒 ( pO) ;
短句来源
     And then,pCI-neo-GPI-G250 was transfected to the RenCa cell successfully.
     结果:构建的重组质粒pCI-neo-GPI-G250经测序正确;
短句来源
     (3) PCI interface technology .
     (3)PCI接口技术。
短句来源
     Summary of PCI Bus
     PCI总线概述
短句来源
     ,Neochonetes gigantea sp., n.
     ,Neo-chonetes gigantea sp. n.
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  pcineo
We have constructed a bicistronic vector, pCINeo/IRES-GFP, which utilises a red-shifted variant of Green Fluorescent Protein as an in vivo cell marker.
      
It is concluded that the bicistronic pCINeo/IRES-GFP vector provides an efficient and non-invasive way of identifying cells which express ion channels after transfection.
      
The expression of the different domains of a complex serine protease, recombinant Factor C of the horseshoe crab, Carcinoscorpius rotundicauda, was achieved using pCIneo and pEGFP-N1 vectors.
      
In an in vitro coupled transcription-translation system, aberrant products were generated from pCDNAI/CrFC21, as opposed to pCIneo/CrFC21 which gave rise to the expected polypeptide.
      
COS-1 cells transfected with pCIneo/CrFC21 expressed Factor C-related proteins which differ from those produced in the yeast, Pichia pastoris.
      
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bjectives To explore that COS7 cells efficiently transfected with human thrombopoietin (hTPO) cDNA from fetal liver and hTPO potentially treated carboplatininduced thrombocytopenia in mice. Methods Using molecular cloning technique, expression vectors PCIneo under the control of SV40 early. Applying liprofectin mediating method, recombinant plasmids PCIneohTPO were transfected into COS7 cells in addition to effect of G418. After hTPO cDNA and mRNA in colony COS7 cells were respectively...

bjectives To explore that COS7 cells efficiently transfected with human thrombopoietin (hTPO) cDNA from fetal liver and hTPO potentially treated carboplatininduced thrombocytopenia in mice. Methods Using molecular cloning technique, expression vectors PCIneo under the control of SV40 early. Applying liprofectin mediating method, recombinant plasmids PCIneohTPO were transfected into COS7 cells in addition to effect of G418. After hTPO cDNA and mRNA in colony COS7 cells were respectively identified by PCRSouthern and dot blot, hTPO was analysed with sandwich ELISA and administrated intraperitoneally to mice with thrombocytopenia. Results COS7 cells transfected with constructed PCIneohTPO expressed and secreted hTPO up to 48.28 ng/ml in the supernatant. We kinetically observed that the number of megakaryocytecolonyforming unit (CFUMK) in bone marrow enhanced to threefold (P<0.01), especially the number of small CFUMK, accompanied by an increased mean megakaryocyte volume (P<0.01) and circulating platelets returned to a normal level (80±26, ×109/L, P<0.01) in peripheral blood. Conclusion Recombinant PCIneohTPO can efficiently be transfected into COS7 cells and hTPO is an effective cytokine for treating carboplatininduced thrombocytopenia in mice.

目的探讨将人血小板生成素(hTPO)cDNA转染COS-7细胞中及表达产物对实验性小鼠血小板减少症(卡铂诱导)的治疗作用。方法以哺乳动物表达载体PCIneo为运载体,构建受控于SV40早期启动子hTPOcDNA的表达载体,利用脂质体介导转染技术,将重组PCIneo-hTPO表达载体转染COS-7细胞中,经G418作用后,对挑选阳性克隆COS-7细胞中hTPOcDNA和mRNA分别进行PCR扩增Southernblot检测和斑点杂交鉴定;对转染COS-7细胞培养上清进行hTPOELISA夹心法检测;应用hTPOcDNA转染的COS-7细胞表达产物对实验性小鼠血小板减少症进行腹腔注射。结果hTPOcDNA不仅可在COS-7细胞中获得表达,且分泌到细胞培养上清中,最高表达量为48.28ng/ml。实验组小鼠第10天,骨髓巨核细胞集落形成单位(CFU-MK)集落数增加近3倍(P<0.01)。骨髓CFU-MK以小集落数为主,巨核细胞体积增大(P<0.01);外周血中血小板数保持在(80±26)×109/L,P<0.01。结论hTPOcDNA转染COS-7细胞中获得有效地表达,且表达的hTPO对实验性小鼠血小板减少症具?

he fulllength 1.1 kb thrombopoietin(TPO) cDNA from fetal liver was cloned into the expressing vector PCIneo to construct the recombinant plasmid PCIneoTPO. COS7 cells were grown to log phase and transfected using LipofectinTM reagent. TPO cDNA of COS7 cells transfected was analyzed with PCR amplificationSouthern blot, mRNA expressed with Dot blot and supernatant with TPO ELISA. The expressing TPO stimulating megakaryocytes in mice in vitro was observed. The results showed that TPO secreted...

he fulllength 1.1 kb thrombopoietin(TPO) cDNA from fetal liver was cloned into the expressing vector PCIneo to construct the recombinant plasmid PCIneoTPO. COS7 cells were grown to log phase and transfected using LipofectinTM reagent. TPO cDNA of COS7 cells transfected was analyzed with PCR amplificationSouthern blot, mRNA expressed with Dot blot and supernatant with TPO ELISA. The expressing TPO stimulating megakaryocytes in mice in vitro was observed. The results showed that TPO secreted from COS7 cells transfected could increase the significantly fourfold number of megakaryocyte colony forming unit(CFUMK) and enhance the size of MK to 4210±6.70 μm(P<001). CFUMK reached peaking level after day 8, then continued for 2 days, and decreased after day 10. These data demonstrate that COS7 cells tranfected with the construct PCIneoTPO by mediated LipofectinTM may successfully express TPO in the supernant liquid which promotes megakaryocytopoiesis.

将克隆全长1.1kb血小板生成素(TPO)cDNA构建重组PCIneoTPO表达载体,利用LipofectinTM介导转染COS-7细胞中,对阳性克隆COS-7细胞的DNA、mRNA和培养上清分别进行PCR扩增Southernblot、斑点杂交和TPO夹心ELISA检测,观察表达产物对小鼠骨髓巨核细胞的作用。结果表明:全长1.1kbTPOcDNA转染COS-7细胞获得表达,最高表达量可达48.28U/ml,其产物使骨髓巨核细胞集落形成单位(CFU-MK)集落数增加4倍,巨核细胞增大至42.10±6.70μm(P<0.01);动态观察骨髓CFU-MK集落数于实验第8天达最高峰,约持续2d,第10天后开始下降。提示TPOcDNA转染COS-7细胞表达产物对骨髓巨核细胞生成有刺激活性

Objective To study the effects of radiation inducible gene on the regulation of the expression of hematopoietic growth factor and the restoration of hematopoiesis.Methods Human GM-CSF cDNA and FLT3 Ligand(FL) were linked together with IRES and inserted into the expression vector pCI-Egr-1 which was constructed with the substitution of CMV promotor in pCIneo with Egr-1 promotor (Egr-FG). The vector was transferred into human bone marrow stromal cell line HFCL with lipofectinTM. The mRNA expression of...

Objective To study the effects of radiation inducible gene on the regulation of the expression of hematopoietic growth factor and the restoration of hematopoiesis.Methods Human GM-CSF cDNA and FLT3 Ligand(FL) were linked together with IRES and inserted into the expression vector pCI-Egr-1 which was constructed with the substitution of CMV promotor in pCIneo with Egr-1 promotor (Egr-FG). The vector was transferred into human bone marrow stromal cell line HFCL with lipofectinTM. The mRNA expression of the target gene in the cells were identified with RT-PCR. The amount of FL and GM-CSF in the supernatant of HFCL-EFG cell culture was determined with ELISA. CD34+cells isolated with MACS were cultured in the medium containing the supernatant of HFCL/EFG to observe its effect on the proliferation of CD34+cells.Results The gene expression vector (Egr-FG) initiated with Egr-1 regulation sequence was successfully constructed. It was confirmed that there were expression of exogenous FL and GM-CSF in HFCL-EFG cells. After 16 hours of 60Co irradiation, the amount of FL and GM-CSF in the serum-free supernatant of HFCL/EFG cell culture was increased respectively(P< 0.001).It was found that after irradiation, the supernatant of HFCL/EFG culture significantly increased its effect on the proliferation of CD34+cells.Conclusion These in vitro data provide an experimental basis for the in vivo use of gene therapy of hematopoietic growth foctor gene regulated by Egr-1 to protect hematopoiesis from ionizing radiation.

目的探索辐射诱导基因调控元件启动的造血生长因子表达及其对造血恢复的作用。方法该实验将携带Egr 1调控序列启动的FLT3配基(FL)和GM CSF双顺反子表达载体(Egr FG)导入骨髓基质细胞系HFCL(称为HFCL/EFG)。采用RT PCR、ELISA及细胞增殖法观察FL和GM CSFcDNA在转染细胞中的表达及保护造血作用。结果构建了Egr 1调控序列启动的双顺反子基因表达载体(Egr FG) ;在HFCL/E FG细胞中证实有外源性FL和GM CSF基因的整合和表达 ,在辐照后16h的HFCL/EFG细胞培养上清液中 ,FL和GM CSF含量较照射前明显增高(P<0.01) ;同时证实辐射后HFCL/EFG培养上清液对造血祖细胞有明显扩增作用 ;共培养的骨髓有核细胞和HFCL/EFG经照射后7d计数非粘附细胞 ,HFCL/EFG组较对照组明显增加(P<0.05)。结论辐射诱导基因Egr 1调控序列启动的造血生长因子基因表达载体在辐射后表达明显增高 ,在体外对辐射后的造血细胞具有保护作用。

 
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