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1.
通过分析比较 2个螺旋藻物种 7个品系胞外和胞内核酸酶活性 ,确定了抑制胞外和胞内核酸酶活性的方法。用液体培养基清洗两次可消除胞外核酸酶活性 ,添加 EDTA(2 m mol L-1)可显著地抑制胞内核酸酶活性。高温 (6 5℃ )处理也能显著抑制胞内核酸酶活性  相似文献   

2.
建立螺旋藻转基因体系初报   总被引:9,自引:1,他引:8  
从钝顶螺旋藻单细胞克隆的制备、重组平台的 克隆、同源重组表达质粒的构建、质粒的电激转化和报告基因的表达等方面对螺旋藻转基因 表达系统进行了研究,初步建立起螺旋藻转基因体系。用次氯酸钠溶液处理获得无菌培养系 ;用钠、钙离子混合液处理,获得了可再生的螺旋藻单细胞。用含EDTA的培养基预培养和用 培养基洗涤可分别去除胞外核酸酶活性和抑制胞内核酸酶活性。以氯霉素乙酰转移酶基因替 换大肠杆菌表达质粒pBV220的抗性选择标记,同时插入系列同源重组片段和萤火虫荧光素酶 基因,构建了同源重组表达载体pBVCS01-04L。电激转化螺旋藻S6-4品系单 细胞,获得了具有稳定氯霉素抗性的培养系,并用SDS-聚丙烯酰胺凝胶电泳证实了报告基 因的表达。  相似文献   

3.
胞内限制性内切酶降解外源DNA,是钝顶螺旋藻/节旋藻(Spirulina/Arthrospira platensis)遗传转化的难点之一。通过用EDTA螯合Mg2 抑制限制性内切酶活性的方法,对外源质粒进行了甲基化修饰。结果表明,修饰后的外源质粒可抵抗限制性内切酶3h的降解,有利于钝顶螺旋藻的遗传转化。  相似文献   

4.
利用啤酒废水养殖螺旋藻研究   总被引:5,自引:0,他引:5  
利用啤酒废水养殖极大螺旋藻(Spirulina marima),研究了废水成分、氮源、藻密度和光照等培养条件对藻生长和蛋白质含量的影响。结果表明,曝气处理的啤酒废水养殖的螺旋藻,相对生长率与CFFRI培养基的几乎一致.蛋白质含量第6天最高,为0.2886g/g干质量,小于CFFRI培养基养殖的。实验确定曝气处理废水养藻的最佳条件是用NaOH调废水pH、藻初始密度取53.8mg/L、光照在1000~10000 1x范围,添加尿素或碳酸氢钠或曝气8h/d。经PSB处理的啤酒废水养殖的螺旋藻,蛋白质为0.4825g/g干质量,与CFFRI培养基养殖的相近。用光合细菌(PSB)处理的废水养藻应控制废水pH为7.0且废水与PSB的体积比为3:1。  相似文献   

5.
钝顶螺旋藻部分原生质体及单细胞的制备与培养   总被引:21,自引:0,他引:21  
于1992年1月-1994年4月,进行超声波处理方法制备钝顶螺旋藻部分原生质体以作为基因工程受体,以及制备单细胞用于固体平板克隆化培养的研究。研究结果表明:超声波以20kHz频率、15W功率作用30s,可使藻丝体断裂成15.0±1.6个细胞长度;延长作用时间,至2-6个细胞长度时,细胞壁结构遭到破坏,形成部分原生质体;继续作用,可形成少量原生质体和大量单细胞。断裂藻丝体、部分原生质体、单细胞以及原生质体均可涂布于固体培养基上再生或生长。以一定密度涂布单细胞与原生质体,能够形成彼此分开的单个克隆,可用于筛选及遗传分析。本文提供了一种节省溶菌酶的制备螺旋藻透性体的方法,超声波作用利于外源基因的导入,而涂布培养利于进一步的筛选和形成克隆。  相似文献   

6.
环境因子对鳗弧菌生长和胞外蛋白酶表达的影响   总被引:2,自引:1,他引:1       下载免费PDF全文
鳗弧菌(Vibrio anguillarum)是一种水产动物的重要致病菌,鳗弧菌分泌的胞外蛋白酶是该菌的致病因子之一.研究了环境因子对鳗弧菌W-1生长及胞外蛋白酶表达的影响.结果表明,鳗弧菌的最适生长温度为28℃,菌体生长量在21 h达到最高,胞外蛋白酶的活力在24 h最高.在37℃培养时,菌体生长和酶活力显著降低;添加葡萄糖、蔗糖、麦芽糖等碳源物质均能显著促进菌体的生长,但对于胞外蛋白酶表达有明显抑制作用;1 mmol/L EGTA对茵体的生长和胞外蛋白酶表达没有明显影响,而1 mmol/L的EDTA强烈抑制鳗弧菌的生长及其胞外蛋白酶的表达.添加不同浓度的氨苄青霉素对菌体的生长及其胞外蛋白酶表达有明显的抑制作用.  相似文献   

7.
用改进的超声波-溶菌酶法制备钝顶螺旋藻原生质球。钝顶螺旋藻(Spirulina platensis)经超声波破碎后,用1.5 mol/L NaCl的Zarrouk培养基洗去细胞外壁的胶质鞘,然后进行酶解。酶解条件为:pH7.2的0.2 mol/L磷酸钾缓冲液,0.8 mol/L KCl,0.5%溶菌酶,28~30℃水浴震荡酶解5~7 h,获得40%以上有活力的钝顶螺旋藻原生质球。  相似文献   

8.
在不同温度和光强下培养极大螺旋藻438(Spirulina maxim438),得到其积累多糖规律为:不利于螺旋藻生长的温度、光照条件,会促进胞外多糖EPS(extracellular polysaccharide,EPS)的分泌,而螺旋藻最适生长条件(30℃,5000lx)与胞内多糖IPS(intracellular polysaccharide,IPS)积累的条件一致。MTT(四甲基偶氮唑盐)法体外实验证明IPS和EPS对人宫颈癌Hela细胞有抑制作用,最佳作用浓度分别为80μg/mL,100μg/mL。Caspase-3试剂盒检测经两种多糖作用后Hela细胞Caspase-3酶的变化,结果显示:处理组比对照组Caspase-3酶活性有明显提高,预示着多糖抑制肿瘤细胞的机制可能是通过Caspase途径,引起细胞的凋亡。  相似文献   

9.
以南极硅藻GJ01(Berkeleya rutilans)为对象,分别采用紫外、可见和荧光分光光度法,研究了营养条件等因子对GJ01生长和胞内谷胱甘肽含量的影响,并对培养的GJ01细胞合成谷胱甘肽的条件进行了优化。结果表明,在单因子影响条件下,1.2g/L的碳酸氢钠对南极硅藻GJ01的生长最好,且最有利于谷胱甘肽的积累;0.8mmol/L氯化钙对GJ01的生长最为有利,而0.2mmol/L氯化钙对谷胱甘肽的产生是适宜的;维生素B_(12)和生物素对GJ01胞内谷胱甘肽的产生影响较小。正交试验表明,GJ01细胞产生谷胱甘肽的最优条件是:光照周期为16h/8h、NaHCO_3为1.6g/L、Ca~(2 )为0.4mmol/L、Cys为10mmol/L。该结果为利用南极硅藻GJ01生产谷胱甘肽提供了参考和依据。  相似文献   

10.
在培养基中添加亚硒酸钠、硫酸钾培养钝顶螺旋藻,研究硒、硫不同克分子比(Se:S)对钝顶螺旋藻有机化硒的影响,用2,3-二氨基萘荧光分光光度法测定藻体中总硒、有机硒、4价硒及6价硒含量,分析藻体中硒的形态,价态构成。结果显示,一定范围内Se:S对钝顶螺旋藻产率影响不大,但硒浓度为3.80mmol/L、Se:S=0.812时螺旋藻产量、总硒及有机硒含量均最高,富硒钝顶螺旋藻中以有机硒为主,无机硒以4价硒为主。  相似文献   

11.
Coastal inundation associated with extreme sea levels is the main factor which leads to the loss of life and property whenever a severe tropical cyclonic storm hits the Indian coasts. The Andhra and Orissa coasts are most vulnerable for coastal inundation due to extreme rise in sea levels associated with tropical cyclones. Loss of life may be minimized if extreme sea levels and associated coastal flooding is predicted well in advance. Keeping this in view, location specific coastal inundation models are developed and applied for the Andhra and Orissa coasts of India. Several numerical experiments are carried out using the data of past severe cyclones that struck these regions. The simulated inland inundation distances are found to be in general agreement with the reported flooding.  相似文献   

12.
This study tests the hypothesis that the in situ void ratio of surficial marine sediments may be predicted from shear wave velocity-depth data with a reliability equal to that of other methods currently available. Shear wave velocity is fundamentally controlled by the number of grain-to-grain contacts per unit volume of material and by the effective stress across those contacts. In this study, three previously established empirical formulae are used to predict void ratio from velocity-depth data. Field data were acquired along a transect off the northern Californian coast across which water depth increased from 35 to 70 m and seafloor sediment type varied from sand to silty-sand, respectively. A towed seafloor sled device was used to collect shear wave refraction data, and a marked, systematic decrease in velocity was observed along the line, ranging from 35-70 m/s for the coarse, near-shore material to 25-40 m/s for the finer, offshore deposits. Void ratios predicted from these velocities were compared with data measured directly from box-core samples. Of the formulae used for prediction, two agree remarkably well with the control data. Both predicted and control values increase from 0.6-0.8 for the sandy material to 1.1-1.5 for the silty-sand. Thus, this study does not disprove the hypothesis set and demonstrates the potential of field shear wave velocity-depth data as a means of delineating spatial variation in void ratio for surficial marine sediments in a remote, nondestructive manner.  相似文献   

13.
Grain size and water content in box-core sediments from the Clarion-Clipperton fracture zone (C-C zone) in the northeast equatorial Pacific were analyzed in detail to understand the downcore variations across a hiatus between Quaternary and Tertiary layers. Grain-size distributions in the topmost core sediments show two modes: a coarse mode (peaked at 50 μm) and a fine mode (at 2-25 μm). The coarse mode disappears gradually with depth accompanied by the dissolution of siliceous fossil tests, whereas the fine mode coarsens due to the formation of authigenic minerals. Water content increases abruptly across a color boundary between an upper pale brown layer and a lower dark brown layer that is the hiatus between Quaternary and Tertiary layers. Abundant smectites and microvoid molds, which are created by the prolonged fossil dissolution in the underlying sediment, are attributed for the abrupt downcore variation of water content. Overall variations in grain size and water content in the topmost core sediments in the western C-C zone are possibly constrained by the dissolution of biogenic siliceous fossils. Variations in geotechnical properties related to these changes must be considered in the design of nodule collectors.  相似文献   

14.
This article concerns an interrelation between the sea levels and the western boundary flow near a tectonic boundary in a local zone in the Northwestern Pacific. In this zone, sea level variations at stations located on the coast facing the Pacific are studied to find the interrelation between variations of the Kurosio flow as an index of the distance of the flow axis off a specific coast. The result is discussed after data processing of the monthly means of the sea levels, and a notice is taken of variations caused by active crustal upheavals during a seismic event, a local earthquake.  相似文献   

15.
Particulate samples were collected from the Changjiang river system during a flood period, in May 1997, and POC, stable isotope and lipids associated with particles were examined. Results showed the decrease (0.84% ~ 1.88%) of organic carbon content from the upper reaches to the estuary.δ13C values of particulate organic carbon was in the range of -24.9×10-3 to -26.6×10-3, which were close to the isotopic signature of continental C3 vegetation. Total particulate n-alkanes concentrations varied from 1.4 to 10.1μg/dm3,or from 23.7 to 107μg/g of total suspended matter. Fatty acids were present in all the samples, from 1.4 to 5.4μg/dm3, with saturated and unsaturated straight-chain and branched compounds in the carbon number range from C12 to C30. Both δ13C and the ratio of carbon content to nitrogen content indicate the predominance of terrestrial inputs (soil organic matter) among the particles. The biomarker approach has been used to identify the relative portion of terrigenous and autochthonous fraction in the particulate samples. The distribution of fatty acids suggests a striking phytoplanktonic and microbial signal in most particle samples. The terrestrial alkanes are used to estimate the contribution of terrestrial inputs along the mainstream.  相似文献   

16.
Current trends of development of satellite derived bathymetry (SDB) models rely on applying calibration techniques including analytical approaches, neuro-fuzzy systems, regression optimization and others. In most of the cases, the SDB models are calibrated and verified for test sites, that provide favorable conditions for the remote derivation of bathymetry such as high water clarity, homogenous bottom type, low amount of sediment in the water and other factors. In this paper, a novel 3-dimensional geographical weighted regression (3GWR) SDB technique is presented, it binds together methods already presented in other studies, the geographically weighted local regression (GWR) model, with depth dependent inverse optimization. The proposed SDB model was calibrated and verified on a relatively difficult test site of the South Baltic near-shore areas with the use of multispectral observations acquired by a recently launched Sentinel-2 satellite observation system. By conducted experiments, it was shown that the proposed SDB model is capable of obtaining satisfactory results of RMSE ranging from 0.88 to 1.23[m] depending on the observation and can derive bathymetry for depths up to 12m. It was also shown, that the proposed approach may be used operationally, for instance, in the continuous assessment of temporal bathymetry changes, for areas important in the context of ensuring local maritime safety.  相似文献   

17.
As a part of the environmental impact assessment studies, geotechnical properties of sediments were determined in the Central Indian Basin. The undrained shear strength and index properties of the siliceous sediments were determined on 20 box cores of uniform dimension collected from various locations in five preselected sites. The maximum core length encountered was 41 cm and most of the sediments were siliceous oozes consisting of radiolarian or diatomaceous tests. The shear strength measurements revealed that surface sediments deposited in recent times (0-10 cm) have a shear strength of 0-1 kPa; this value increases with depth, reaching 10 kPa at 40 cm deep. Older sediments have greater strength because of compaction. Water content varies in the wide range of 312-577% and decreases with depth. The clay minerals such as smectite and illite are dominant and show some control over water content. Wet density, specific gravity, and porosity do not indicate any notable variation with depth, thereby indicating a uniform, slow rate of sedimentation. The average porosity of sediments is 90.2%, specific gravity 2.18, and wet bulk density 1.12 g/cm 3 . Sediments exhibit medium to high plasticity characteristics, with the average plasticity index varying between 105% and 136%. Preliminary studies on postdisturbance samples showed an increase in natural water content and a decrease in undrained shear strength of sediments in the top 10- to 15-cm layer.  相似文献   

18.
Several major improvements to an existing method for calibrating satellite altimeters using tide gauge data are described. The calibration is in the sense of monitoring and correcting temporal drift in the altimetric time series, which is essential in efforts to use the altimetric data for especially demanding applications. Examples include the determination of the rate of change of global mean sea level and the study of the relatively subtle, but climatically important, decadal variations in basin scale sea levels. The improvements are to the method described by Mitchum (1998a), and the modifications are of two basic types. First, since the method depends on the cancellation of true ocean signals by differencing the altimetric data from the tide gauge sea level time series, improvements are made that produce a more complete removal of the ocean signals that comprise the noise for the altimetric drift estimation problem. Second, a major error source in the tide gauge data, namely land motion, is explicitly addressed and corrections are developed that incorporate space-based geodetic data (continuous GPS and DORIS measurements). The long-term solution, having such geodetic measurements available at all the tide gauges, is not yet a reality, so an interim solution is developed. The improved method is applied to the TOPEX altimetric data. The Side A data (August 1992?February 1999) are found to have a linear drift component of 0.55 + / 0.39 mm/yr, but there is also a significant quadratic component to the drift that is presently unexplained. The TOPEX Side B altimeter is estimated to be biased by 7.0 + / 0.7 mm relative to the Side A altimeter based on an analysis of the first 350 days of Side B data.  相似文献   

19.
Extreme sea levels associated with severe cyclonic storms are common occurrences along the east coast of India. The coastal districts of Orissa have experienced major surges in the past. The recent Paradip super cyclone is one of the most severe cyclones, causing extensive damage to property and loss of lives. Extreme sea levels are major causes for coastal flooding in this region. Damages can be minimized if the extreme sea levels are forecast well in advance. In the present study, we develop a location specific, fine resolution model for the Orissa coast on the lines similar to that of IIT-D storm surge model (Dube et al. 1994). The model runs on a personal computer. The bathymetry for the model is extracted from very fine resolution naval hydrographic charts for the region extending from the south of Orissa to south of West Bengal. A simple drying scheme has also been included in the model in order to avoid the exposure of land near the coast due to strong negative sea surface elevations. An attempt was made in this study to simulate extreme sea levels along the Orissa coast using the data of past severe cyclones. The model results reported in the present study are in good agreement with available observations or estimates.  相似文献   

20.
The continental margin of northern Sinai and Israel, up to Haifa Bay, is the northeastern limb of the submarine Nile Delta Cone. It is made up predominantly of clastics from the Nile and its predecessors. The continental shelf and coastal plain of Israel are built of a series of shore-parallel ridges composed of carbonate-cemented quartz sandstone (locally named kurkar), a lithification product of windblown sands that were piled up into dunes during the Pleistocene. The drop in global sea level and regression during the last glacial period exposed the continental shelf to subaerial erosion and created a widespread regional erosional unconformity which is expressed as a prominent seismic reflector at the top of the kurkar layers. The subsequent Holocene transgression abraded much of the westernmost kurkar ridges, drowned their cores, and covered the previous lowstand deposits with marine sands, which were in turn covered by a sequence of sub-Recent clayey silts. The Mediterranean coasts of Sinai and Israel are part of the Nile littoral cell. Since the building of the Aswan dams the sand supplied to Israel's coastal system is derived mainly from erosion of the Nile Delta and from sands offshore Egypt that are stirred up by storm waves. The sands are transported by longshore and offshore currents along the coasts of northern Sinai and Israel. Their volume gradually declines northward with distance from their Nile source. The longshore transport terminates in Haifa Bay where some sand is trapped, and the test escapes to deeper water by bottom currents and through submarine canyons, thus denying Nile-derived sand supply to the 40-km-long 'Akko-Rosh Haniqra shelf. The sand balance along Israel's coastal zone is a product of natural processes and human intervention. Losses due to the outgoing longshore transport, seaward escape, and landward wind transport exceed the natural gains from the incoming longshore transport and the abrasion of the coastal cliffs. The deficit is aggravated by the construction of (1) seaward-projecting structures that trap sands on the upstream side and (2) offshore detached breakwaters that trap sands between themselves and the coast. The negative sand balance is manifested by the removal of sand from the seabed and the consequent exposure of archaeological remains that were hitherto protected by it. The sediments that escape seaward from the longshore transport system form a 2.5- to 4-km-wide sandy apron adjacent to the shore that extends to where the water is 30 - 40 m deep. The apron's slope (0.5 - 0.8) is steeper than the theoretical equilibrium slope for the median grain-size diameter in this zone (0.1 - 0.3 mm). The beach sands and the apron's surficial sands are well sorted. Their grain size decreases with distance from shore, from 0.2 - 0.3 mm nearshore to 0.11 - 0.16 mm by the drowned ridge. The coarse-grained fraction consists of skeletal debris (commonly 5 - 12% carbonate matter) and wave-milled kurkar grains (locally named zifzif). In deeper water, the basal sands underlying the fine-grained sediment cover consist of 1- to 30-cm layers whose composition ranges from silty sands to various types of sands (fine, medium, coarse, and gravelly) to zifzif. For the most part, they contain large amounts of skeletal debris (20 - 60%) and small fragments of kurkar. Two types of kurkar rock were encountered offshore: a well-sorted, fine- to medium-grained (0.074 - 0.300 mm) lithified dune sand with variable amounts of carbonate cement, ranging from hard rock of low permeability to loose sand; and a porous sandstone made up predominantly of algal grains and skeletal debris (calcarenite).  相似文献   

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