65 resultados para RT-QPCR

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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Zebrafish has been generally considered as an excellent model in case of drug screening, disease model establishment, and vertebrate embryonic development study. In this work, the ability of human cytomegalovirus immediate early promoter (CMV promoter)-driven short hairpin RNA (shRNA) expression vector to induce shRNA against VEGF gene in zebrafish was tested, and its effect on vascular development was assed, too. Using RT-qPCR, blood vessel staining, and in situ hybridization, we confirmed certain transcriptional activity and down regulation of gene expression by the vector. In situ hybridization analysis indicated selective inhibition of NRP1 expression in the VEGF gene loss of function model, which might imply in turn that VEGF could not only activate endothelial cells directly but also could contribute to stimulating angiogenesis in vivo by a mechanism that involved up-regulation of its cognate receptor expression in zebrafish. This contributed to a better understanding of molecular mechanisms of cardiovascular development. The system improved the success rate in making inducible knockdown and widened the possibilities for better therapeutic targets in zebrafish.

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A variety of N-acetyl-o-aryl-1,2-didehydroethylamines were synthesized by direct reduction-acetylation of beta-aryl-nitroolefins and assayed as HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTIs) for the first time. Compound 7a exhibited a TI v

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应用RT -PCR方法 ,通过用不同浓度的甲基汞对Hep2B细胞株进行不同时间长短的处理 ,检测其调钙质的mRNA表达水平 ,发现微量的甲基汞处理就可以引起细胞株细胞损伤 ,同时抑制调钙质mRNA的表达 ,进一步证实了在肝脏损伤的病理状态下调钙质基因的表达减少 ,从而为检测环境中甲基汞的含量提供了一个可能的分子生物学手段 .

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A rapid, sensitive and highly specific detection method for Aquareovirus based on reverse-transcription polymerase chain reaction (RT-PCR) was developed. Based on multiple sequence alignment of the cloned sequences of a local isolates, the Threadfin reovirus (TFV) and Guppy reovirus (GPV) with Grass carp reovirus (GCRV), a pair of degenerate primers was selected carefully and synthesized. Using this primer combination, only one specific product, approximately 450 bp in length was obtained when RT-PCR was carried out using the genomic double-stranded RNA (dsRNA) of TFV, GPV and GCRV. Similar results were also obtained when Chum salmon reovirus (CSRV) and Striped bass reovirus (SBRV) dsRNA were used as templates. No products were observed when nucleic acids other than the dsRNA of the aquareoviruses described above were used as RT-PCR templates. This technique could detect not only TFV but also GPV and GCRV in low titer virus-infected cell cultured cells. Furthermore, this method has also been shown to be able to diagnose GPV-infected guppy (Poecilia reticulata) that exhibit clinical symptoms as well as GPV-carrier guppy. Collectively, these results showed that the RT-PCR amplification method using specific degenerate primers described below is very useful for rapid and accurate detection of a variety of aquareovirus strains isolated from different host species and origin. (C) 2004 Elsevier B.V. All rights reserved.

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The ability of the Evpatoria RT-70 radar complex to perform research on space debris was investigated in four trial experiments during 2001-2003. The echo-signals of 25 objects at geostationary, highly elliptical and medium-altitude orbits were recorded on magnetic tapes at radio telescopes in Russia, Italy, China and Poland. The multi-antenna system configuration gives potential to supplement the classic radar data with precise angular observations using the technique of Very Long Baseline Interferometry. The first stage of such processing was fulfilled by the correlator in N. Novgorod, Russia. The cross-correlation of transmitted and received signals was obtained for the 11 objects on the Evpatoria-Bear Lakes, Evpatoria-Urumqi and Evpatoria-Noto baselines. This activity also promoted developing the optical observations of geostationary objects, conducted for the improvement of the radar target ephemerides. (C) 2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

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为调查葡萄植株携带葡萄卷叶病毒(GLRaV)的情况,以带毒和非带毒指示植物“品丽珠”为试验材料提取总RNA,并以此总RNA为模板进行RT-PCR扩增,建立了GLRaV的RT-PCR检测体系,并对GLRaV cDNA序列进行测定,结果成功地检测到了GLRaV。经多次试验证明,该检测结果准确可靠,可以用于GLRaV的检测。

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The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been integrated into the genome of gametophytes of L. japonica, and the expression product showed the expected bioactivity, implying the proper post-transcript modification in haploid gametophyte.

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The InAsxSb1-x films were grown on (100) GaSb substrates by liquid-phase epitaxy, and their structural, electrical, and optical properties were investigated. The high-resolution x-ray diffraction results reveal that the single crystalline InAsxSb1-x films with a midrange composition are epitaxially grown on the GaSb substrates. Temperature dependence of the Hall mobility was theoretically modeled by considering several predominant scattering mechanisms. The results indicate that ionized impurity and dislocation scatterings dominate at low temperatures, while polar optical phonon scattering is important at room temperature (RT). Furthermore, the InAsxSb1-x films with the higher As composition exhibit the better crystalline quality and the higher mobility. The InAs0.35Sb0.65 film exhibits a Hall mobility of 4.62x10(4) cm(2) V-1 s(-1). The cutoff wavelength of photoresponse is extended to about 12 mu m with a maximum responsivity of 0.21 V/W at RT, showing great potential for RT long-wavelength infrared detection. (C) 2008 American Institute of Physics. [DOI: 10.1063/1.2989116]

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The growth of InAsxSb1-x films on (100) GaSb substrates by liquid-phase epitaxy (LPE) has been investigated and epitaxial InAs0.3Sb0.7 films with InAs0.9Sb0.09 buffer layers have been successfully obtained. The low X-ray rocking curve FHWM values of InAs0.3Sb0.7 layer shows the high quality of crystal-orientation structure. Hall measurements show that the highest electron mobility in the samples obtained is 2.9 x 10(4) cm(2) V-1 s(-1) and the carrier density is 2.78 x 10(16)cm(-3) at room temperature (RT). The In As0.3Sb0.7 films grown on (10 0) GaSb substrates exhibit excellent optical performance with a cut-off wavelength of 12 mu m. (c) 2007 Elsevier B.V. All rights reserved.

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本文给出一种用激波管研究材料烧蚀的方法。这一方法的基础是对烧蚀气相产物进行全样色谱分析,求得气相产物的总量,最后直接得到样品的烧蚀速率。用此方法对泰氟隆材料进行研究求得在高温下聚四氟乙烯的表面烧蚀速率为1.6×10~-2exp(-18400/RT)mol?Cm~-2?s~-1,(1400K

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We report on a diode-pumped, cryogenic and room temperature operation of a Tm,Ho:YAlO3 (c-cut) laser. In a temperature of 77 K, an optical-optical conversion efficiency of 27% and a slope efficiency of 29% were achieved with the maximum continuous-wave (CW) output power of 5.0 W at 2.13 mu m. Acousto-optic switched operation was performed at pulse repetition frequency (PRF) from 1 kHz to 10 kHz, the highest pulse energy of 3.3 mJ in a pulse duration of 40 ns was obtained. In room temperature (RT), the maximum CW power of Tm,Ho:YAlO3 laser was 160 mW with a slope efficiency of 11% corresponding to the absorbed pump power. (C) 2008 Optical Society of America.

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Aluminum nitride (AlN) films were prepared on gamma-LiAlO2 substrates by radio frequency (rf) magnetron sputtering. The influence of substrate temperature (T-s) and nitrogen (N-2) concentration on film growth was investigated. The X-ray diffraction (XRD) results reveal that highly c-axis oriented AlN films can be obtained in the temperature range from room temperature (RT) to 300 degrees C. A smoother surface and a crystalline quality decrease with increasing N-2 concentration have been observed by XRD and atomic force microscopy (AFM) for the films deposited at lower substrate temperature. On the contrary, the degradation of the surface smoothness and the higher crystalline quality can be observed for the films deposited at a higher substrate temperature with N-2-rich ambient. The growth mechanism which leads to different crystalline quality of the films is discussed. (C) 2008 Elsevier B.V. All rights reserved.