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以杂色鲍(Haliotis diversicolor Reeve)为研究对象,通过密度梯度离心方法纯化得到高纯度血蓝蛋白,以其作为抗原皮下注射免疫新西兰大白兔,从而获得高效价的兔源多克隆抗血清。进一步通过Protein A抗原亲和纯化的方法对该抗血清纯化,最终获得效价更高、检测特异性更好的血蓝蛋白多克隆抗体。应用该抗体进行Western检测发现,鲍血淋巴中存在着多样的血蓝蛋白衍生产物;进一步结合质谱技术对其中35 kDa条带进行鉴定发现,其来源于血蓝蛋白I型亚基的H结构域。 相似文献
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泥蚶肌肉组织蛋白质双向电泳体系的建立 总被引：4，自引：0，他引：4
本研究建立了泥蚶肌肉组织蛋白质的双向电泳体系.提取泥蚶肌肉组织全蛋白,进行了双向电泳分离的初步尝试,并对各个关键因素和环节进行了优化比较.优化的裂解液配方是8mol/dm3 Urea,1.5mol/dm3 Thioruea,4% CHAPS,10mmol/dm3 Tris,2mmol/dm3 TBP,0.2%Bio-Lyte3/10ampholyte,1mmol/dm3 PMSF以及1mmol/dm3 EDTA.使用pH值为4～7的IPG胶条进行被动水化上样,等电聚焦采用缓慢升压模式,电泳参数设置和染色方法等根据Bio-rad公司推荐的进行调整.最终获得了较满意的双向电泳图谱,具有较高的分辨率和重复性,成功建立了泥蚶肌肉组织蛋白质双向电泳分离体系. 相似文献
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为了研究中国鲎血液质量的监测方法,实验选择9只中国鲎检测其血液中铜含量、血细胞数量、血浆蛋白含量、血淋巴蛋白含量等指标,并进行相关性分析。结果表明,中国鲎血液中铜含量与血细胞数相关性r=0.917,与血浆蛋白含量相关性r=0.995,与血淋巴蛋白含量相关性r=0.983,均呈现极显著相关性。中国鲎血液中铜含量与血细胞数的相关关系为Y1=6.14+0.86X(R2=0.840),与血浆蛋白含量的相关关系为Y2=2.08+0.78X(R2=0.989),与血淋巴蛋白含量的相关关系为Y3=1.87+0.62X(R2=0.966)。中国鲎血液中铜含量不但能够直接反映血蓝蛋白的含量,而且可以反映血细胞数、血浆蛋白含量、血淋巴蛋白含量。因此,以中国鲎血液中铜含量作为监测血液质量的指标,可以推动中国鲎可持续采血技术的发展与完善,对中国鲎的利用与保护具有重要意义。 相似文献
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本文对实验室前期研制的抗中国明对虾(Fenneropenaeus chinensis)血蓝蛋白单克隆抗体进行特性分析,并以该单抗为第一抗体,采用石蜡切片、免疫组化方法,研究了血蓝蛋白在中国明对虾鳃、心、胃、中肠、淋巴器官、卵巢和肝胰脏等组织器官中的分布特点.结果表明:血蓝蛋白在鳃丝小叶边缘的微血腔、心肌束间隙、淋巴器官的淋巴腔中以及肝小管中阳性信号较强;在卵巢卵母细胞之间的血窦、胃内壁环肌层结缔组织的血窦和中肠结缔组织血窦中有阳性信号.结论认为,中国明对虾血蓝蛋白阳性信号大多来自血淋巴而非实质组织,且在血淋巴含量多、血流量大的部位含量丰富,在某些血淋巴无法到达的部位(肝小管内壁)也有血蓝蛋白分布,血蓝蛋白含量的这种组织特异性与各组织器官的功能有着密切的联系. 相似文献
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卵黄蛋白原和血蓝蛋白是凡纳滨对虾(Litopenaeusvannamei)卵巢和/或肝胰腺合成的重要蛋白。利用半定量和荧光定量PCR技术分别检测了亲虾卵巢不同发育期卵黄蛋白原和血蓝蛋白mRNA在卵巢和肝胰腺的相对表达量。结果显示,卵黄蛋白原mRNA在卵巢和肝胰腺中均有表达,其相对表达量随着卵巢发育持续增加,在第五期达到最高,在恢复期迅速下降。根据卵巢和肝胰腺质量的综合计算,凡纳滨对虾的卵黄蛋白中,卵巢的贡献率占主导地位,约占80%,而肝胰腺仅为卵巢的四分之一。血蓝蛋白mRNA主要在肝胰腺中表达,随着卵巢的发育,表达量呈增加趋势,至第三期达到高峰,其后稍有下降,但仍然维持在较高水平,在第六期恢复到初始水平。血蓝蛋白是否直接或间接参与了卵黄发生和卵巢发育有待进一步探讨研究。 相似文献
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注射病原菌对凡纳滨对虾血蓝蛋白合成、酚氧化酶活力的影响 总被引：1，自引：0，他引：1
研究注射病原菌对凡纳滨对虾血蓝蛋白合成、酚氧化酶活力的影响。结果表明:注射哈维氏弧菌对凡纳滨对虾血淋巴中血蓝蛋白含量、肝胰脏内血蓝蛋白mRNA和p75、p77亚基表达以及血淋巴、血细胞、血蓝蛋白酚氧化酶活力的影响显著(P<0.05),而对照组无显著变化。血蓝蛋白含量和p75、p77亚基表达在0～24 h内呈峰值变化,其中血蓝蛋白含量在6 h内逐渐下降、且6 h时达到最低值,而p75、p77亚基表达在3 h内无明显变化,然后均表现为逐渐上升趋势,各指标在12 h时达到最大值,而且亚基表达与注射哈维氏弧菌浓度呈正相关,24 h后各指标趋于稳定,与对照组无显著差异。在注射哈维氏弧菌短时间内血蓝蛋白mRNA表达呈迅速上升趋势,其中注射6×106cells/mL处理组mRNA表达在6h内呈峰值变化,3 h时达到最大值,6～48 h内保持稳定,但仍明显高于对照组水平(P<0.05),而注射6×105cells/mL处理组mRNA表达量虽然增加,但只在6 h达到最大值时与对照组差异显著(P<0.05),其余时间均与对照组差异不显著。血淋巴、血细胞和血蓝蛋白酚氧化酶活力分别在12 h,24 h内呈峰值变化,其中血淋巴、血蓝蛋... 相似文献
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采用SDS聚丙烯酰胺凝胶电泳方法,对锯缘青蟹肌肉中可溶性蛋白质进行分离,以过敏患者阳性血清和正常人阴性血清作对照,通过Western blot鉴定出分子量为14.3kDa的蛋白为锯缘青蟹的特异性过敏原.然后用Sephadex G75凝胶过滤层析和DEAE-52离子交换层析纯化出该过敏原,再经MALDI-TOF/TOF-MS检测,利用Blastz在线分析.结果表明,锯缘青蟹特异性过敏原为血蓝蛋白亚基,经Clustal W分析该蛋白的多肽片段序列与可口美青蟹、邓杰内斯蟹、加州龙虾、艾氏真蟹的血蓝蛋白亚基序列有很高的同源性,经Prosite数据库检索后发现肽段1和肽段2有酪蛋白激酶Ⅱ磷酸化位点和蛋白激酶C磷酸化位点,是过敏原参与机体反应的重要活性位点. 相似文献
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为研究中国明对虾(Fenneropenaeus chinensis)血蓝蛋白C末端(FcHC-C)的抗菌功能,将血蓝蛋白基因FcHC的2个C末端基因片段连接到毕赤酵母表达载体pPIC9K中,构建酵母表达载体pPIC9K/FcHC-C。该载体经SalI酶切后,采用PEG法转化毕赤酵母(Pichia pastoris GS115)。转化子经过PCR鉴定后,阳性克隆通过含有G418的YPD平板筛选,获得高拷贝重组子。重组毕赤酵母利用甲醇诱导表达目的基因。经Tricine-SDS-PAGE和Western blot分析结果表明,利用酵母工程菌成功表达了血蓝蛋白C末端片段(rFcHC-C1和rFcHC-C2)。抑菌活性鉴定实验结果显示,重组蛋白rFcHC-C1和rFcHC-C2作为阴离子抗菌肽具有抗真菌和抗细菌的活性。 相似文献
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Chandralata Raghukumar G. Sheelu P. A. Loka Bharathi Shanta Nair C. Mohandass 《Marine Georesources & Geotechnology》2001,19(1):1-16
In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance. 相似文献
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Derek V. Ellis 《Marine Georesources & Geotechnology》2001,19(1):51-63
This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested. 相似文献
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G. Parthiban 《Marine Georesources & Geotechnology》2000,18(3):223-235
Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement. 相似文献
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Arne Schnack-Friedrichsen Angela M. Davis Jim D. Bennell Dei G. Huws 《Marine Georesources & Geotechnology》2001,19(4):221-243
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths. 相似文献
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Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification
An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm. 相似文献
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海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。 相似文献
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The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments. 相似文献
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Shigehisa Nakamura 《Marine Geodesy》2000,23(1):55-61
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone. 相似文献
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