适用于小麦叶片总蛋白分析的双向电泳技术

双向电泳技术越来越成为植物蛋白质组研究中的关键技术,样品制备、固相预制胶条水化、等电聚焦及SDS—PAGE都是影响双向电泳结果的重要步骤,本研究以晋麦47号小麦叶片为研究材料,在样品除盐、预制胶条水化方式、第二向SDS—PAGE电压选择方面进行了探索与优化,通过比较双向电泳图谱,建立适用于小麦叶片总蛋白质分析的双向电泳技术体系．结果表明：样品经多步除盐并加长除盐时间后进行双向电泳,蛋白能较好地被分离,2-DE图谱蛋白点较多,分辨率较高;IPG胶条先后经被动水化与主动水化比仅主动水化的2-DE图谱蛋白点损失少;第二向SDS—PAGE中电压选择120V较200V的2．DE图谱蛋白点拖尾少,纹理现象少．     

油菜双向电泳技术体系的优化

为建立一套可以较好地分离油菜不同器官(花蕾、化药、叶片)的双向电泳技术体系,对IPG胶条的pH范围、凝胶染色的方法以及蛋白质的上样量进行优化.结果表明,以pH=4~7,17cm的IPG胶条进行双向电泳,胶体考马斯亮蓝染色,蛋白质上样量为800μg时,油菜的花蕾、化药以及叶片蛋白质均可得到很好的分离,获得的2-DE凝胶图谱清晰.为检验优化后的双向电泳技术体系与质谱的兼容性,选取10个蛋白质点进行肽质量指纹图谱分析,发现这些蛋白质点经质谱分析后均可得到成功鉴定.表明该分离体系与质谱兼容性较好,蛋白质点鉴定成功率较高.     

耐辐射菌Micrococcus luteus SC1204总蛋白双向电泳体系的建立

以耐辐射藤黄微球菌Micrococcus luteus SC1204为研究对象,建立总蛋白双向电泳体系.结果表明,采用液氮研磨-酚/超高速离心法提取总蛋白,裂解液Ⅱ(8 mol/L尿素、2mol/L硫脲、60mmol/L DTT、4%CHAPS、40mmol/L Tris、1%pH 3~10NL IPG buffer、0.002%BPB)溶解蛋白,使用24cm、pH 4~7的IPG胶条,上样量250μg及等电聚焦7h(56000Vh),可获得满意分辨率的双向电泳图谱,适用于后续的质谱及差异蛋白质组分析.     

金钗石斛花芽蛋白质提取及其双向电泳体系优化

通过对蛋白质提取方法、水化方式、等点聚焦程序、IPG胶条选择等方面的优化，建立金钗石斛花芽蛋白质双向电泳体系.结果表明，采用酚抽提法提取蛋白，样品与水化液1∶〖KG-*2/3〗40复溶；被动水化，等点聚焦程序B2；选用24 cm pH 3～10NL 胶条和12% 的凝胶；采用考马斯亮蓝G-250染色，获得了可识别1 422个蛋白质斑点的双向电泳图谱，且重复性好、分辨率高，为进一步开展金钗石斛花芽在蛋白质组学水平上的分析提供了技术支持.     

青藏高原有机牦牛肉背最长肌双向电泳体系的建立

为了建立青藏高原有机牦牛肉背最长肌双向电泳(2-DE)体系,本研究对肌肉样品的处理、蛋白质的提取以及等电聚焦方式等进行了研究。结果表明:(1)用超声波裂解肌肉蛋白质的检出率及蛋白分离效果明显优于用液氮研磨;(2)用直接裂解法(7M尿素,2M硫脲,4%CHAPS(w/v),0.5%两性电解质p H 3~10,100 mmol/L DTT(w/v),60 mmol/L Tris,20μL蛋白酶抑制剂)提取肌肉组织全蛋白获得的蛋白质量、凝胶图谱斑点数目、匹配率和图像分辨率最好,尤其适合提取小分子蛋白;(3)采用间隔式等电聚焦方式,蛋白质的分离效果明显优于普通升压方式,且横条纹较少,蛋白点数量增多。结论:本研究建立的2-DE体系,能提高有机牦牛背最长肌总蛋白双向电泳的分辨率和重复性,为后续进行有机牦牛肉品质评价的蛋白质组学研究,更好地解析青藏高原有机牦牛肉品质形成的遗传调控机制奠定了基础。     

大鼠骨组织蛋白质组样品提取方法的建立

蛋白质组学是目前研究的重要热点．该拟建立大鼠骨组织蛋白质样品提取的最佳方法．用于双向电泳分离．为进一步进行蛋白组学分析奠定基础．实验对3种不同方法制备的样本进行同一条件的二维电泳(2-DE)图谱．比较2-DE的结果．结果发现，丙酮沉淀法最佳．这些结果提示．只有结合骨组织成分特征与双向电泳的具体要求．选择恰当的样品提取液．采用合理的提取步骤，获得蛋白质样品，才能得到清晰高分辨率的电泳图谱，满足蛋白质组学分析需要。     

缺铁逆境胁迫下水稻叶蛋白质组的双向电泳分析

水稻幼苗经缺铁胁迫诱导处理1、3、5d后，用酚法和TCA／丙酮法提取叶的可溶性蛋白，进行双向电泳(2-DE)分析．结果显示：(1)三种缺铁胁迫的可溶性蛋白，在不同的pH范围中2-DE图谱上的比较．使用pH3～10宽范围、线性(L)的IPG胶条，经电泳分离后利用Z3图象分析软件，可在SDS-PAGE凝胶上检测到450个左右的蛋白点，酸性蛋白占89％．选用pH4～7窄范围、线性(L)的IPG胶条，经电泳后可在SDS-PAGE凝胶上检测到600个左右的蛋白点，其中缺铁诱导上调表达的有29个点，减弱表达的有1个点，诱导特异表达的有5个点．(2)不同方法提取可溶性蛋白的差异．酚法提取的蛋白再溶性好，纯度较高．TCA／丙酮法简单易操作，蛋白损耗少，在2-DE图象上．碱性端显示的蛋白点较多，但此法的缺点是再溶性差，对2-DE成功分离蛋白有影响．     

大麻蛋白质组的双向电泳技术建立

蛋白组学研究中最关键步骤是样品制备,为得到高质量的双向电泳蛋白图谱,作者通过对TCA/丙酮法对大麻茎、叶进行蛋白样品制备、双向电泳技术流程进行优化,建立了适于大麻茎、叶的双向电泳技术体系.优化双向电泳体系为:裂解液在传统配方中加入Tris-HCl(p H=8.8),以Triton X-100代替CHAPS,蛋白裂解后加入4倍体积100%预冷丙酮等均可有效去除各种非蛋白杂质;IPG胶条p H=4~7,24 cm,考染上样量800μg/胶条可得到背景清晰、分辨率高的双向电泳图谱.大麻茎中可分辨的蛋白点为1 027±12,叶中为905±8.该体系能同时适用于大麻茎、叶,并能满足下一步蛋白质谱测序分析的要求,可为大麻蛋白组学的发展提供技术支持.     

甘蔗叶片蛋白质组双向电泳技术优化

为了建立适合甘蔗叶片蛋白质组研究的双向电泳技术,对甘蔗叶蛋白质的溶解方法、IEF电泳、上样量等关键步骤进行了优化.结果表明:裂解液中有2 mmol/L的硫脲才能更充分地溶解蛋白;上样量1.0 mg时得到质量较好的凝胶图谱;甘蔗叶片蛋白质主要分布在pH4～7范围.通过对甘蔗叶片蛋白双向电泳技术的优化,提高了双向电泳图谱分...     

优化海兔肝蛋白质组提取与分离技术

为了提高海兔肝脏蛋白质组的提取及研究效率，本文描述和比较了3种提取海兔肝脏蛋白质组的方法：裂解液浸泡-超速离心，丙酮-TCA沉淀-裂解液浸泡，裂解液浸泡-丙酮-TCA-裂解液溶解．然后采用常规SDS-PAGE双向电泳对提取出的蛋白质进行检验，用Melanie 4 Trial软件分析电泳图谱，通过对蛋白质分辨率和斑点总数的比较，选出一种更适合海兔肝蛋白质的提取方法．结果表明采用丙酮-TCA沉淀-裂解液浸泡法提取海兔肝蛋白质所得的双向电泳图谱不论从蛋白质斑点分辨率还是总数上都优于其他两种方法，更适合提取分离海兔肝脏蛋白质．     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

海南岛地体及其毗邻陆缘晚中生代—新生代古地磁研究和构造演化

本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。     

Exciton Seebeck in Molecular Systems

Various applications relevant to the exciton dynamics,such as the organic solar cell,the large-area organic light-emitting diodes and the thermoelectricity,are operating under temperature gradient.The potential abnormal behavior of the exicton dynamics driven by the temperature difference may affect the efficiency and performance of the corresponding devices.In the above situations,the exciton dynamics under temperature difference is mixed with     

Achieving Unusual States of Matter Under High Pressure

The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to     

《Chinese Science Bulletin》SUBJECT INDEX TO VOLUME 59(2014)

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An on-line scaling method for improving scalability of a database cluster

The explosive growth of the Internet and database applications has driven database to be more scalable and available, and able to support on-line scaling without interrupting service. To support more client's queries without downtime and degrading the response time, more nodes have to be scaled up while the database is running. This paper presents the overview of scalable and available database that satisfies the above characteristics. And we propose a novel on-line scaling method. Our method improves the existing on-line scaling method for fast response time and higher throughputs. Our proposed method reduces unnecessary network use, i.e. , we decrease the number of data copy by reusing the backup data. Also, our on-line scaling operation can be processed parallel by selecting adequate nodes as new node. Our performance study shows that our method results in significant reduction in data copy time.     

An improved R-tree based on childnode''s probability

R-Tree is a good structure for spatial searching. But in this indexing structure,either the sequence of nodes in the same level or sequence of traveling these nodes when queries are made is random. Since the possibility that the object appears in different MBR which have the same parents node is different, if we make the subnode who has the most possibility be traveled first, the time cost will be decreased in most of the cases. In some case, the possibility of a point belong to a rectangle will shows direct proportion with the size of the rectangle. But this conclusion is based on an assumption that the objects are symmetrically distributing in the area and this assumption is not always coming into existence. Now we found a more direct parameter to scale the possibility and made a little change on the structure of R-tree, to increase the possibility of founding the satisfying answer in the front sub trees. We names this structure probability based arranged R-tree (PBAR-tree).     

Integrating GIS Web services based on mediating architecture

The geographic information service is enabled by the advancements in general Web service technology and the focused efforts of the OGC in defining XML-based Web GIS service. Based on these models, this paper addresses the issue of services chaining,the process of combining or pipelining results from several interoperable GIS Web Services to create a customized solution. This paper presents a mediated chaining architecture in which a specific service takes responsibility for performing the process that describes a service chain. We designed the Spatial Information Process Language (SIPL) for dynamic modeling and describing the service chain, also a prototype of the Spatial Information Process Execution Engine (SIPEE) is implemented for executing processes written in SIPL. Discussion of measures to improve the functionality and performance of such system will be included.     

Tracking data management using topology of moving object database system for moving object constrained trajectory in LBS application

Advances in wireless technologies and positioning technologies and spread of wireless devices, an interest in LBS (Location Based Service) is arising. To provide location based service, tracking data should have been stored in moving object database management system (called MODBMS) with proper policies and managed efficiently. So the methods which acquire the location information at regular time intervals then, store and manage have been studied. In this paper, we suggest tracking data management techniques using topology that is corresponding to the moving path of moving object. In our techniques, we update the MODBMS when moving object arrived at a street intersection or a curved road which is represented as the node in topology and predict the location at past and future with attribute of topology and linear function. In this technique, location data that are corresponding to the node in topology are stored, thus reduce the number of update and amount of data. Also in case predicting the location,because topology are used as well as existing location information, accuracy for prediction is increased than applying linear function or spline function.