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     Inducible expression of LacZ gene in Schwann cells by Adeno-X Tet-On system
     Adeno-X Tet-On系统调控LacZ基因在雪旺细胞中表达的研究
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     Establishment of Conditional Expression of c-myc in Transgenic Mice and Tumor Model by the Tet-On System
     Tet-On系统诱导表达c-myc转基因小鼠及肿瘤模型的建立
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     Effect of conditional expression of esp1 gene by Tet-on system on ploidy in A_549
     Tet-on系统诱导esp1基因表达对A_(549)细胞染色体的影响
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     Establishment of transgenic mice conditional expressing c-myc gene by the Tet-on system
     Tet-on系统诱导表达c-myc转基因小鼠的建立
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     Conditional expression of simian virus 40T antigen by the Tet-on system in CHO
     Tet-on系统诱导猿猴病毒40T在CHO细胞中的表达
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     system).
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     SYSTEM
     系统
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  on system
Research on system architecture of modeling and simulation
      
II International Conference on System Identification and Control Problems
      
II International Conference on System Identification and Control Problems
      
Mathematical modeling of individual and public health based on system models is a vital problem.
      
Based on system analysis of a duel situation of a long-range air fight of aircraft-fighters, the "Attack" and "Defense-with-Attack" problem substitutions are highlighted.
      
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The cDNA of mouse Binlb homolog(mBinlb) protein was subcloned into expression vector of pCDNA3 and transiently expressed in COS-7 cells. The expression of mBinlb in COS-7 cells was identified by RT-PCR.The anti-microbial activity of the transfected cell culture supernatant was confirmed by plating on growth media. Adding a HA tag to the C-terminus of mBinlb (mBinlb-HA)resulted in the same anti-microbial activity as the mBinlb protein. This result indicated that anti-microbial activity was not located at the...

The cDNA of mouse Binlb homolog(mBinlb) protein was subcloned into expression vector of pCDNA3 and transiently expressed in COS-7 cells. The expression of mBinlb in COS-7 cells was identified by RT-PCR.The anti-microbial activity of the transfected cell culture supernatant was confirmed by plating on growth media. Adding a HA tag to the C-terminus of mBinlb (mBinlb-HA)resulted in the same anti-microbial activity as the mBinlb protein. This result indicated that anti-microbial activity was not located at the C terminus of mBinlb. To study the function of mBinlb in vivo, the Tet-on gene expression regulation system was used to construct transgenic mice. Two lines of transgenic mice were obtained, one in which the mBinlb-HA cDNA was under the control of the TRE promoter,and the other with rtTA specifically expressed using the promoter of the mE-RABP gene in the epididymis. Crossing the two transgenic lines will produce inducible over-expression of mBinlb specifically in epididymis. This research will provide a useful mice model to study the function of mBinlb in vivo.

我们在真核细胞中表达了小鼠Binlb基因,证实了它的抗菌活性,并进一步证明了该蛋白C端融合HA抗原决定簇后基本不影响蛋白的生物活性,为开展这一蛋白的转基因小鼠工作提供了较理想的转基因材料。同时我们利用四环素调控的基因表达系统(Tet-on系统)分别建立了附睾专一性表达rtTA的转基因小鼠和由TRE控制的mBinlb-HA的转基因阳性小鼠。这项研究对于我们理解mBinlb在体内的生理和病理作用提供了进一步研究的基础。

Objective To infect Schwann cells by Adeno-X Tet-On system and observe the inducible expression of LacZ gene under the regulation of doxcycline. Methods ①Five to six-day-old Sprague-Dawley rats were sacrificed by decapitation and the sciatic nerves were immediately removed aseptically to get Schwann cells. After the cells had been cultured and purified, those were identified by indirect immunocytochemistry with mouse anti-S100 protein antibody McAb.②Adeno-X Tet-On virus stock and Adeno-X TRE-βgal virus stock...

Objective To infect Schwann cells by Adeno-X Tet-On system and observe the inducible expression of LacZ gene under the regulation of doxcycline. Methods ①Five to six-day-old Sprague-Dawley rats were sacrificed by decapitation and the sciatic nerves were immediately removed aseptically to get Schwann cells. After the cells had been cultured and purified, those were identified by indirect immunocytochemistry with mouse anti-S100 protein antibody McAb.②Adeno-X Tet-On virus stock and Adeno-X TRE-βgal virus stock were amplified by infecting HEK293 cells respectively. When CPE was evident,the viruses were isolatedand viral titer were determined.③Schwann cells were infected with amplified viruses and the data of inducible expression of LacZ gene under the regulation of doxcycline were ploted. Results ①The purity of Schwann cells was more than90% and in a good state. ②The titer of amplified viruses was 1.26×10 9 and1.99×10 9 pfu/ml respectively. Conclusion After Schwann cells were infected with Adeno-X Tet-On virus and Adeno-X TRE-β gal virus, it was found that the concerntration change of Dox had a strong influence on the expression of LacZ gene.

目的 应用Adeno XTet On系统感染雪旺细胞 ,通过四环素衍生物强力霉素 (doxcycline ,Dox)调控LacZ基因产物 (βgal)的表达。方法 ①取生后 5~ 6dSD仔鼠坐骨神经进行雪旺细胞培养 ,并进行形态学及S 10 0蛋白相关抗原免疫组织化学染色鉴定。②将Adeno XTet OnVirusStock与Adeno XTRE βgalVirusStock分别感染HEK2 93细胞 ,收获病毒并作滴度测定。③将扩增后的病毒感染雪旺细胞并绘制Dox调控下βgal诱导表达曲线。结果 ①由体外培养获得的雪旺细胞纯度可达 90 %以上并且状态良好 ,免疫组织化学染色阳性结果明显。②扩增后的病毒滴度较高 ,分别达 1.2 6× 10 9、1.99× 10 9pfu/ml。 结论 将病毒感染雪旺细胞后 ,经检测发现DOX对 βgal表达调控明显

Both tetracycline-controlled transactivator(tTA) and reverse tTA(rtTA) of which tTA activiates transcription in the absence of doxycycline and rtTA in the presence of Dox,respectively,have allowed temporal and spatial control over gene expression in cells and transgenic mice, providing unique genetic models for the study of development process and high-order functions.Simian virus 40T(SV 40T) antigen is a potent oncoprotein that can induce several types of tumors.SV40T antigen acts as a potent growth stimulator...

Both tetracycline-controlled transactivator(tTA) and reverse tTA(rtTA) of which tTA activiates transcription in the absence of doxycycline and rtTA in the presence of Dox,respectively,have allowed temporal and spatial control over gene expression in cells and transgenic mice, providing unique genetic models for the study of development process and high-order functions.Simian virus 40T(SV 40T) antigen is a potent oncoprotein that can induce several types of tumors.SV40T antigen acts as a potent growth stimulator by inactivating p53 and Rb tumor suppressor genes,leading to uncontrolled cellular proliferation and tumor formation.Constructed the tetracycline-induced Simian Virus 40T antigen expression vector in the Tet-on system and expressed it in vitro by the transfection and selection of double-stable cell lines.Report demonstrated value of this double-stable cell lines in studying physiological function of the SV40T antigen.

四环素诱导表达系统(T et-off/T et-on系统)是比较成熟的真核生物基因诱导表达系统之一,具有高效、无毒、严密开/关功能的特点。猿猴病毒40T(SV 40T)是一种病毒癌蛋白,其与肿瘤抑制蛋白p53和R b结合,并使之失活,从而消除它们抑制细胞生长的功能,使细胞分裂加速,形成肿瘤。利用T et-on系统首先稳定筛选获得了表达T et-on系统调节元件rtTA的阳性细胞CHO-pT et-on,再通过稳定筛选又成功得到导入其反应元件的双阳性细胞CHO-pT et-on-pTRE 2-SV 40T-Hyg,经强力霉素诱导表达了目的基因SV 40T,建立了T et-on基因诱导表达系统的细胞诱导表达研究平台。

 
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