共查询到12条相似文献,搜索用时 63 毫秒
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【目的】对Galectin-3蛋白进行纯化,优化诱导相关表达条件,为大量获取罗非鱼Galectin-3蛋白提供方案。【方法】根据NCBI上已公布的罗非鱼(Oreochromismossambicus)Galectin-3基因序列,设计多对带Eco RI和Xhol酶切位点的引物,筛选出良性扩增引物,对罗非鱼cDNA经进行聚合酶链式反应(PCR)扩增、限制性快切酶双酶切、T4连接酶连接等步骤,构建罗非鱼Galectin-3基因的原核表达质粒pGEX-4T-Galectin-3,将重组质粒导入大肠杆菌BL21中,进行Galectin-3重组蛋白的表达。并比较不同的诱导温度、诱导时间、IPTG浓度条件下的表达效果,对Galectin-3蛋白表达最优条件进行筛选。【结果】构建了罗非鱼Galectin-3原核表达质粒,进行Galectin-3重组蛋白后发现37℃下,0.4 mmol/L浓度的IPTG诱导5 h即可诱导Galectin-3重组蛋白高效表达。免疫印迹结果显示,经蛋白纯化柱纯化后的pGEX-4T-Galectin-3重组蛋白可与GST-Tag单克隆抗体发生特异性反应,进而表明表达的重组蛋白是罗非鱼的Galectin-3蛋白。【结论】本研究成功构建了罗非鱼Galectin-3基因的原核表达载体,并确定了Galectin-3融合蛋白最适的表达条件:37℃下,0.4 mmol/L浓度的IPTG诱导5 h。 相似文献
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《中国海洋大学学报(英文版)》2017,(1)
Marine organisms constitute unlimited resource of bioactive substances due to their high biodiversity and represent a valuable source of new compounds. This study optimized the alkali-extraction conditions and antioxidant activities of soluble polysaccharides from the body wall of Sipunculus nudus. The effects of solid-liquid ratio, extraction duration, extraction temperature, and alkali concentration on the yield of S. nudus polysaccharides(SNP) were examined, according to which the optimal combination of extraction parameters was obtained by an orthogonal test. The relative influencing importance of different extraction parameters on the yield of SNP followed the order as solid-liquid ratio extraction temperature alkali concentration extraction duration. The highest extraction yield, 1.98%, was achieved under an optimal extraction condition: temperature 60℃, solid-liquid ratio 1:6 g mL~(-1), duration 5 h, and alkali(Na OH) mass fraction 6%. The in vitro antioxidant activities examination showed that extracted SNP under this optimized condition had strong power in reducing certain hydroxyl and superoxide radical scavenging abilities. The promising results showed that extracted SNP could be a potent natural antioxidant. 相似文献
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锆石U-Pb定年是近些年来发展起来的一种很成熟的定年方法,快速、简单。该次研究在ArF193nm激光剥蚀器和iCAPQ电感耦合等离子体质谱仪之间设计添加了外置匀化器,从而使激光剥蚀出来的气溶胶可进行混匀后再被载气载入等离子体质谱仪,这不仅提高了等离子体质谱仪的信号稳定性,同时大大提高了锆石测试区域的精密度和准确度,为线状剥蚀扫描进行不同成分域定年的开展提供了便利条件。利用通过加载外置器皿的联接条件对新疆天山造山带北部的二长花岗岩11k88进行线状剥蚀取样U-Pb年龄测试,显示加载最优外置器皿后,线状剥蚀取样的测试结果与前人进行SHRIMP U-Pb年龄测定结果在误差范围内一致。表明加载人工设计的匀化器进行激光器和等离子体质谱仪之间的联接优化,可进行线状浅剥蚀取样方式的锆石U-Pb年龄和微量元素测定,这将拓展LA-ICP-MS在地质年代学研究中的应用。 相似文献
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Hf是一种高场强元素,在地壳中的丰度较低,往往以类质同象的形式进入锆石中。以氟铪酸钾(K_2HfF_6)络合物为研究对象,观察它们在富水热液中的水解程度,进而研究Hf元素在地质过程中的迁移行为。结果表明:K_2HfF_6络合物在热液条件下能发生一定程度的水解反应,总体趋势为络合物的初始质量浓度越低且反应的温度越高,其水解程度越剧烈,络合物的稳定性也就越低;获得了高温高压条件下K_2HfF_6络合物的累积水解平衡常数。同时运用实验模拟的方法,根据提取液中Hf元素含量来推测矿物结晶时富氟流体中Hf元素含量,若以K_2HfF_6络合物最高配位的离子团形迁移时,Hf与F元素之间的质量浓度比值为1.6,据此可以计算成矿流体所需的最小F元素含量。 相似文献
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中国地质科学今后一个时期的发展趋势和重点 总被引:10,自引:0,他引:10
近代地质学从诞生之日起就担负起了两方面的使命,一是了解地球,揭示地球作为一个自然实体的内在规律性:二是以获得的地球知识为人类社会服务。按照“研究与发展”(R&D)的划分.前者属于基础研究.后者属于应用研究。通常认为,地质学是以基础研究与应用研究紧密联系为特色的。到了20世纪,特别是其后期,这两项使命仍然是最根本的,但其广度和深度都有了惊人的拓展。 相似文献
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《中国海洋大学学报(英文版)》2017,(5)
Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose(8.0%), skim milk(12.0%), sodium citrate(2.0%), serum(12.0%) and PVP(2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of Na Cl was 10.0, 30.0 and between 5.0 and 10.0 g L~(-1) for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at-80℃ or-40℃ for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate(79.63%–82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy(SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda. 相似文献
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精细地质研究现状及发展趋势 总被引:5,自引:2,他引:5
水驱采油后期,油层的含水率越来越高,然而大量的可动剩余油却滞留于地下。要提高这部分油层的采收率,必须加强对储层微观非均质性的认识。精细地质研究作为一种方法,在这种背景下,便得到了快速的发展和应用。旨在对其做一个阶段性的小结,从储集层砂体几何形态、内部结构以及孔、渗空间变化特征出发,概述了精细地质研究的内容为细化开发单元、成因单元砂体的连续性和连通性描述、砂体内部建筑结构单元的划分、流动单元的研究、表外储层研究、地质建模等。并提出了今后研究发展的主要方向。为油田的可持续发展提供技术支持。 相似文献
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复杂断块构造具有断层复杂,小断层广泛发育,断层识别难度大,储层分布及连通性复杂,地震成像不清、资料品质较差等特征。对于此类问题,常规地震资料无法满足断层解释精度要求。提出了一种新的断层EDF解释性处理技术,将地震资料处理与解释融为一体,以提高资料的信噪比和断层的识别能力。断层EDF解释性处理技术是利用扩散滤波和中值倾角滤波相结合的断层增强滤波方法来提高资料的信噪比,突出断层和地层的接触关系,再在滤波后的地震资料数据体上进行分频处理,选择最佳成像频带,突出识别不同规模的断层。通过在涠洲A油田的应用,取得了良好的效果。 相似文献