165 resultados para silver-stained denaturing PAGE


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生物多样性科学(BiodiversityScience)的国际规划提出生物多样性对生态系统功能的影响是整个研究计划五大核心的核心.生物多样性包括遗传、物种和生态系统三个水平,其中遗传多样性是其它两个水平多样性的基础和最终来源.该文在实验室多年研究毛乌素沙地柠条遗传多样性的基础上,分别从表型(生理生化)、蛋白质、同工酶以及遗传型(rDNA)水平探讨中间锦鸡儿根瘤菌的遗传多样性,并模拟沙地生境,建立人工共生体系,以期发现最有效的共生伙伴关系,这不仅有得提高毛乌素地区农牧业产量,更重要的是在当今沙尘暴肆虐的情况下,发挥柠条防风固沙的能力具有现实意义. 1.毛乌素沙地中间锦鸡儿根瘤菌遗传多样性(1)全细胞可溶性蛋白质谱将供试中间锦鸡儿根瘤菌菌株分为两大类群,其中硬梁覆沙地菌株GH72不同于来自沙丘顶部和底部的菌株,而且中间锦鸡儿根瘤菌独立于参比菌株。酯酶同工酶谱分析表明,中间锦鸡儿根瘤菌与参比菌株仅存在一个等位酶位点差异,其余等位点与参菌株共享,因此,酯酶同工酶反映出中间锦鸡儿根瘤菌的异质性。(2)16SrDNA部分序列与16S-23S rDNA IGS结果表明,所有供试菌株扩增产物均较前人报道的分子量偏高。经16S rDNA PCR-RFLP分析,中间锦鸡儿根瘤菌共形成12种基因型,表现出丰富的遗传多样性,其中属于基因型2的菌株占42.4%。代表菌株GH33 16S rDNA全序列结果显示,与已知的快生型根瘤菌同源性在95%以上。(3)中间锦鸡儿根瘤菌生理生化反应特性B.T.B实验证明所有中间锦鸡儿根瘤菌均产酸,符合快生型根瘤菌的特征.唯一碳源测试显示,95%中间锦鸡儿根瘤菌不利用淀粉,33%菌株不利用乳糖,对其他测试碳源不具有选择性。检洲在不同盐离子浓度、不同酸性梯度以及不同温度条件下菌株生长状况,发现毛乌素沙地中间锦鸡儿根瘤菌具极强的耐盐性.53.8%的菌株可以在9%NaCl的YMA培养基生长.75%的菌株在pH4.O和pHl0,0 环境中仍能生长,66.7%菌株在60℃处理1 0min后仍具有生活力。体现出对于干旱沙地的适应。 2.不同实验共生系统中植物和根瘤菌对生态系统功能的影响14株根瘤菌分与三个柠条种(小叶锦鸡儿,中间锦鸡儿和柠条锦鸡儿)回接,用土壤上覆沙模拟毛乌索沙地景观生态条件,以多石砾贫瘠土壤为对照,比较不同基因型柠条与根瘤菌人工共生体的长和结瘤与生境的关系,初步证明根瘤菌很可能是该生态系统的关键种。寄主植物与共生根瘤菌的遗传多样性对生态系统功能的影响与生态环境有关。实验还表明,选择适当的共生组合对于防治沙漠化有很大潜力。3.银染变性聚丙烯酰胺凝胶电泳检测RAPD遗传模式以85株小钻杨F2代为材料,用本实验室改良的银染变性聚丙烯酰胺凝胶电泳法检测RAPD遗传模式。结果表明,仅用9个引物共扩增到399个位点,其中98个位点表现为多态性,卡方测验显示,79个多态位点符合经典的孟德尔遗传(3:1),占多态位点80.6%。这种改良的检测RAPD标记的方法必将推动RAPD标汜构建连锁图谱的进程。

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OBJECTIVE: To determine whether rDNA of Giardia lamblia forms a nucleolus organizer region (NOR)-like structure and is in a very primitive state. STUDY DESIGN: G lamblia was used as the experimental animal, with Euglena gracilis as the control. The distribution was demonstrated indirectly by the modified Ag-I silver technique, which can specifically indicate the NOR under both light and electron microscopes. RESULTS: In the ultrathin sections of silver-stained Euglena cells, all the silver grains were concentrated in the fibrosa of the nucleolus, while no grains found in the cytoplasm, nucleoplasm, condensed chromosomes or pars granulosa of the nucleus. In the silver-stained Giardia cells, no nucleolus was found; a few silver grains were scattered in the nucleus but were not concentrated in any specific region. CONCLUSION: The distribution of silver grains in G lamblia showed that the transcription of rDNA occurs inside the nucleus, though no nucleolus is present. It is possible that chromosomes are in a very primitive state in diplomonad cells; as each chromosome has few prRNA genes, the transcription is independent of a nucleolus. These results imply that the rDNA of Giardia does not form a NOR-like structure and seems to represent a very primitive state in the evolution of the nucleolus.

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Fea's tree rat (Chiromyscus chiropus) is a very rare species which there are only a few specimens in the world. The chromosomes of two male specimens, collected from Xishuanbanna, Yunnan, are analysed by several banding technique (G-, C-bands, as well as Ag-staining). The diploid chromosome number is 22, and autosomes comprise 5 pairs of metacentrics, 2 pairs of subacrocentrics, and 3 pairs of acrocentrics. The X chromosome is a acrocentric, and Y is a micro-chromosome, almost a point, which could be a marker chromosome of the species and the genus. The centromeric C-bands are very faint, and C-bands of Nos. 1, 2, 9 and Y chromosome are negative. Only one pair Ag-NORs was found on No. 10 in the silver-stained karyotype. The relationship between morphologic and chromosomal features was discussed, and C-banded karyotype evolutionary trend has also been discussed. Moreover, the conventional karyotype of Niviventer confucianus was described.

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R-phycoerythrin was isolated and purified from a red alga, Polysiphonia urceolata Grev, using Streamline column combined with ion-exchange chromatography or hydroxyapatite chromatography. The purity of R-phycoerythrin isolated by Streamline column was up to 1.66 and the yield of R-phycoerythrin could be as high as 0.68 mg/g frozen P. urceolata. All the eluates from Streamline column were divided into two equivalent parts, respectively. One part was pumped into the ion-exchange column loaded with Q-Sepharose and the other was applied to the adsorption column loaded with hydroxyapatite. The purities of R-phycoerythrin purified using these two methods were both up to 3.26, more than 3.2 the commonly accepted criterion. The yield of purified R-phycoerythrin from the ion-exchange chromatography was 0.40 mg/g frozen P. urceolata and that from the hydroxyapatite chromatography could reach 0.34 mg/g frozen P. urceolata. The purified protein had three absorption peaks at 498, 535, and 565 nm and displayed a fluorescence maximum at 580 nm, which was consistent with the typical spectrum of R-phycoerythrin. The purified R-PE was also identified with electrophoresis. Only one single protein band appeared on native-PAGE with silver staining. SDS-PAGE demonstrated the presence of one 20 kDa major subunit, and one low intensity band corresponding to 33 kDa subunit. The results indicate that using the expanded bed adsorption combined with ion-exchange chromatography or hydroxyapatite chromatography, R-phycoerythrin can be purified from frozen P. urceolata on large scale. (c) 2006 Elsevier Inc. All rights reserved.

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We investigated the effect of cerium oxide on the precipitation of Ag nanoparticles in silicate glass via a femtosecond laser irradiation and successive annealing. Absorption spectra show that Ce3+ ions may absorb part of the laser energy via multiphoton absorption and release free electrons, resulting in an increase of the concentration of Ag atoms and a decrease of the concentration of hole-trapped color centers, which influence precipitation of the Ag nanoparticles. In addition, we found that the formed Ag-0 may reduce Ce4+ ions to Ce3+ ions during the annealing process, which inhibits the growth of the Ag nanoparticles.

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The authors report the investigation of filament and supercontinuum generation by focusing a femtosecond laser beam into water doped with silver nanoparticles. The silver nanoparticles enhance the nonlinear optical response of water, leading to broadening of supercontinuum spectra in self-focused femtosecond filaments. During the propagation of the supercontinuum light in the filament, the silver nanoparticles preferentially scatter the short-wavelength light near the plasmon resonant wavelength peak, followed by the scattering of the long-wavelength light. Thus, a side view of the filament shows a full-color spectrum in the visible range, which is herein called "rainbow filament." (c) 2007 American Institute of Physics.

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Silver nanowires in large quantities can be obtained through a simple method in the absence of a surfactant or polymer and without addition of external seeding nanocrystallites. A plausible mechanism was proposed to elucidate the formation mechanism of silver nanowires based on TEM studies.

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Recent theoretical and experimental results suggested that the silver superlens could be constructed through controlling silver thin film thickness and preparation conditions, and applied in subdiffraction-limited optical imaging and optical lithography. In this work, we report another significant application of silver superlens-ultrahigh density optical data storage. With the silver superlens the subdiffraction-limited pit arrays on an optical disk are dynamically read out and the carrier-to-noise ratio can reach 25 dB for the thin film thickness of 46 nm. The readout laser power and readout velocity have little effect on the carrier-to-noise ratio. Additionally, in our experiment the silver thin film thickness needs to be controlled in the range from 20 to 80 nm.

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Fast moving arrays of periodic sub-diffraction-limit pits were dynamically read out via a silver thin film. The mechanism of the dynamic readout is analysed and discussed in detail, both experimentally and theoretically. The analysis and experiment show that, in the course of readout, surface plasmons can be excited at the silver/air interface by the focused laser beam and amplified by the silver thin film. The surface plasmons are transmitted into the substrate/silver interface with a large enhancement. The surface waves at the substrate/silver interface are scattered by the sinusoidal pits of sub-diffraction-limit size. The scattered waves are collected by a converging lens and guided into the detector for the readout.

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We report on space-selective co-precipitation of silver and gold nanoparticles in Ag+, Au3+ co-doped silicate glasses by irradiation of femtosecond laser pulses and subsequent annealing at high temperatures. The color of the irradiated area in the glass sample changed from yellow to red with the increase of the annealing temperature. The effects of average laser power and annealing temperature on precipitation of the nanoparticles were investigated. A reasonable mechanism was proposed to explain the observed phenomena. (c) 2006 Elsevier Ltd. All rights reserved.

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Coatings with layer structures of Ag/glass, Ag/Cr/glass, and Ag/Cr-Ag/Cr/glass deposited with magnetron sputtering are investigated. The results indicate that the performance for reflectance, hardness, adherence, and humidity durability of the silver coatings with Ag/Cr/glass and Ag/Cr-Ag/Cr/glass structures are better than pure silver film. In addition, the silver coatings with an Ag/Cr-Ag/glass structure present more advantages than that of the Ag/Cr/glass. Reasons are analyzed accordingly. (C) 2004 Society of Photo-Optical Instrumentation Engineers.

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A series of silver films with different thickness were prepared under identical conditions by direct current magnetron sputtering. The optical properties of the silver films were measured using spectrophotometric techniques and the optical constants were calculated from reflection and transmission measurements made at near normal incidence. The results show that the optical properties and constants are affected by films' thickness. Below the critical thickness of 17 nm at which Ag film forms a continuous film, the optical properties and constants vary significantly as the thickness of films increases and then tends to a stable value which is reached at 41 nm. X-ray diffraction measurements were carried out to examine the structure and stress evolution of the Ag films as a function of films' thickness. It was found that the interplanar distance of (111) orientation decreases when the film thickness increases and tends to be close to that of bulk material. The compressive strains also decrease with increasing thickness. (C) 2007 Published by Elsevier B.V.