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1.
Isolation of viral coat protein mutants with altered assembly and aggregation properties 总被引:1,自引:0,他引:1 下载免费PDF全文
A method was developed to screen bacteria for synthesis of mutant proteins with altered assembly and solubility properties using bacteriophage MS2 coat protein as a model self-associating protein. Colonies expressing coat protein from a plasmid were covered with an agarose overlay under conditions that caused the lysis of some of the cells in each colony. The proteins thus liberated diffused through the overlay at rates depending on their molecular sizes. After transfer of the proteins to a nitrocellulose membrane, probing with coat protein-specific antiserum revealed spots whose sizes and intensities were related to the aggregation state of coat protein. The method was employed in the isolation of assembly defective mutants and to find soluble variants of an aggregation-prone coat protein mutant. 相似文献
2.
Chen Chen Raymond Dagnino Jr. Charles Q. Huang James R. McCarthy Dimitri E. Grigoriadis 《Bioorganic & medicinal chemistry letters》2001,11(24):3165-3168
Cyclizations of alkylhydrazines with N-acyl-S-methylisothioureas, readily synthesized from acyl chlorides, sodium thioisocyanate, dialkylamines then methyl iodide in a one-pot reaction, gave 1-alkyl-3-dialkylamino-5-phenyltriazoles 7 as major products. The regioisomers were assigned through the use of NOE NMR experiments. While bearing a N-bis(cyclopropyl)methyl-N-propylamino group, this series of compounds shows very good binding affinity on the human CRF1 receptor. Among them, 1-methyl-3-[N-bis(cyclopropyl)methyl-N-propylamino]-5-(2,4-dichlorophenyl)-1H-[1,2,4]triazole 7a had the best binding affinity for the CRF1 receptor (Ki=9 nM). 相似文献
3.
Lina Long Douglas R. McCabe M. Eileen Dolan 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,731(2):128
A highly sensitive and selective method for determining 8-oxoguanine in plasma and urine was developed by high-performance liquid chromatography with electrochemical detection. The compound was separated by gradient elution on a C18 reversed-phase column with a mobile phase of acetonitrile and 0.1 M sodium acetate, pH 5.2. 8-Hydroxy-2′-deoxyguanosine was used as internal standard. 8-Oxoguanine was detected electrochemically by setting the potential to +300 mV vs. Pd reference. The sensitivity of the assay was 22 ng/ml with a signal-to-noise ratio of 7:1. The within-day relative standard deviations for 8-oxoguanine quality control samples with concentrations of 3340, 1340 and 84 ng/ml were 3.6, 4.3 and 5.7% for plasma, and 4.1, 4.6 and 6.2% for urine, respectively. The day-to-day relative standard deviations for the same samples were 3.8, 6.8 and 7.1% for plasma, and 3.9, 7.0 and 7.9% for urine, respectively. The method is designed to study the pharmacokinetics and metabolic fate of O6-benzylguanine in a phase I clinical trial. Previously, O6-benzyl-8-oxoguanine was identified as the primary metabolite of O6-benzylguanine in humans. We now demonstrate that 8-oxoguanine is a further metabolite of O6-benzylguanine. 相似文献
4.
Lina L. E. Massone 《Biological cybernetics》1994,71(4):293-305
This paper presents a neural-network-based system that can generate and control movements of the eyes. It was inspired by a number of experimental observations on the saccadic and gaze systems of monkeys and cats. Because of the generality of the approach undertaken, the system can be regarded as a demonstration of how parallel distributed processing principles, namely learning and attractor dynamics, can be integrated with experimental findings, as well as a biologically inspired controller for a dexterous robotic orientation device. The system is composed of three parts: a dynamic motor map, a push-pull circuitry, and a plant. The dynamics of the motor map is generated by a multi-layer network that was trained to compute a bidimensional temporal-spatial transformation. Simulation results indicate (1) that the system is able to reproduce some of the properties observed in the biological system at the neural and movement levels and (2) that the dynamics of the motor map remains stereotyped even when the motor map is subject to abnormal stimulation patterns. The latter result emphasizes the role of the topographic projection that connects the motor map to the push-pull circuitry in determining the features of the resulting movements. 相似文献
5.
6.
From ligulate flowers of Matricaria chamomilla was isolated a mixture of apigenin 7-O-β-glucoside diacetates, which was shown to be based on (2″, 3″)- and (3″, 4″)-diacetates. 相似文献
7.
Nucleoside permeation in L1210/AM cells is mediated by (a) equilibrative (facilitated diffusion) transporters of two types and by (b) a concentrative Na(+)-dependent transport system of low sensitivity to nitrobenzylthioinosine and dipyridamole, classical inhibitors of equilibrative nucleoside transport. In medium containing 10 microM dipyridamole and 20 microM adenosine, the equilibrative nucleoside transport systems of L1210/AM cells were substantially inhibited and the unimpaired activity of the Na(+)-dependent nucleoside transport system resulted in the cellular accumulation of free adenosine to 86 microM in 5 min, a concentration three times greater than the steady-state levels of adenosine achieved without dipyridamole. Uphill adenosine transport was not observed when extracellular Na+ was replaced by Li+, K+, Cs+, or N-methyl-D-glucammonium ions, or after treatment of the cells with nystatin, a Na+ ionophore. These findings show that concentrative nucleoside transport activity in L1210/AM cells required an inward transmembrane Na+ gradient. Treatment of cells in sodium medium with 2 mM furosemide in the absence or presence of 2 mM ouabain inhibited Na(+)-dependent adenosine transport by 50 and 75%, respectively. However, because treatment of cells with either agent in Na(+)-free medium decreased adenosine transport by only 25%, part of this inhibition may be secondary to the effects of furosemide and ouabain on the ionic content of the cells. Substitution of extracellular Cl- by SO4(-2) or SCN- had no effect on the concentrative influx of adenosine. 相似文献
8.
D Vijayalakshmi L Dagnino J A Belt W P Gati C E Cass A R Paterson 《The Journal of biological chemistry》1992,267(24):16951-16956
Cultured mouse leukemia L1210 cells express the nucleoside-specific membrane transport processes designated es, ei, and cif. The es and ei processes are equilibrative, but may be distinguished by the high sensitivity of the former to 6-[(4-nitrobenzyl)thio]-9-beta-D-ribofuranosylpurine (NBMPR); the cif process is mediated by a Na+/nucleoside cotransporter of low sensitivity to NBMPR. Cells of an ei-deficient clonal line, L1210/MC5-1, were mutagenized, and clones were selected in soft agar medium that contained (i) NBMPR (an inhibitor of es processes), (ii) erythro-9-(2-hydorxy-3-nonyl)adenine (an inhibitor of adenosine deaminase), and (iii) arabinofuranosyladenine (a cytotoxic substrate for the three nucleotide transporters). The selection medium did not allow es activity and selected against cells that expressed the Na(+)-linked cif process. Cells of the L1210/B23.1 clonal isolate were deficient in cif transport activity, and inward fluxes of formycin B, a poorly metabolized analog of inosine, were virtually abolished by NBMPR in these cells. In the mutant cells, nonisotopic formycin B behaved as a countertransport substrate during influx of [3H]formycin B, and inward fluxes of the latter were competitively inhibited by purine and pyrimidine nucleosides. The transport behavior of L1210/B23.1 cells indicates that (i) the mutation/selection procedure impaired or deleted the Na(+)-linked cif process and (ii) es nucleoside transport activity is expressed in the mutant cells. 相似文献
9.
基于遥感的建筑物高度快速提取研究综述 总被引:1,自引:0,他引:1
近年来我国城市化进程不断推进,不仅体现在城市面积上的增长,也体现在建筑物高度的增长。高度增长一方面能尽量克服城市土地资源匮乏的瓶颈,另一方面为优化城市结构及城市功能做出贡献。在城市遥感研究领域,对于城市建筑物高度的提取也成为研究的重点。城市建筑物高度的估计与测量,已成为城市规划和扩张、城市灾害风险预警与评估的重要参数,同时也为数字城市三维模型的建立提供了基础测绘资料。分别基于光学遥感影像、高分辨率SAR(Synthetic Aperture Radar)影像以及光学遥感影像与高分辨率SAR影像的融合三方面,全面阐述城市建筑物高度的提取方法,并比较两类影像在提取建筑物高度的优劣势,通过回顾早年研究方法,逐步引入近年来新的发展趋势。 相似文献
10.
城市化进程中城市热岛景观格局演变的时空特征——以厦门市为例 总被引:4,自引:0,他引:4
热岛效应作为城市化过程中产生的特有环境问题,对其形成和演变规律的研究有助于人们提出有效的应对措施。以厦门市为研究对象,利用1987—2007年等时间间隔、同时相的5景Landsat TM/ETM+遥感影像数据进行地表温度反演,在此基础上使用景观格局指数分析厦门城市热岛景观格局随城市化进程演变的趋势。结果表明:随着厦门城市化进程加深,整个热岛景观在逐渐变得更加破碎化,高等级热岛景观斑块个数、类型面积和个体面积都增大;新的高等级热岛景观斑块都出现在原有高等级斑块附近,致使高等级类型的邻近度增加而各类型之间相互接触的程度也增加;景观总体的聚合度逐渐下降,而高等级热岛景观类型的聚合度则呈上升趋势;景观水平的蔓延度总体呈下降趋势,优势度高的低等级热岛景观所占的比重下降,优势度逐渐降低;多样性指数、均匀度指数总体呈上升趋势,各热岛景观面积在各类型间的分配逐渐趋于均匀;热岛景观斑块的转化方面,在20 a间低等级斑块类型(1、2、3级)向高等级斑块类型(4、5、6级)转化的面积总体上呈增加趋势,而高等级斑块类型向低等级斑块类型转化的面积总体上呈减小趋势,且等级升高的面积明显大于同期等级降低的面积;就高等级热岛景观斑块而言,他们与3级热岛景观斑块间的相互转化最容易发生,远比高等级斑块内部各类型之间的相互转化来得容易,尤其6类和5类的转化是最为困难的热岛景观变化之一;从空间上看,各高等级热岛景观斑块都经历了数量增加、面积扩大、等级升高三个方面的变化,形成了海沧、新阳、杏林、厦门岛西北港口区和机场5个高温组团。利用景观指数分析城市热环境,可探明热岛景观随城市化演变的趋势,并为有效的热岛效应减缓措施提供直接的理论依据。 相似文献