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1.
为了探索核移植克隆动物技术中的细胞融合参数,进行了小鼠2-细胞期胚细胞的电融合试验。直流电场强度为2kV/cm,脉冲过程40μs时,效果较好,可以获得70.5%的融合率和64.5%的发育率,进一步提高电场强度,裂解死亡率也明显提高;采用非电解质溶液作为融合液。  相似文献   

2.
在细胞融合过程中化学聚集电融合法,即用PEG作为聚合剂与电融合法连用,取得明显的效果。其条件是:电压11kV;脉冲宽度5μm;脉冲个数3;脉冲时间59s;温度4℃;PEG浓度35%;处理时间30min;处理温度30℃及pH6.从细胞体积,DNA含量,淀粉糖化能力及葡萄糖发酵能力等方面说明,所获得的新型酵母菌株是两亲本的融合株。  相似文献   

3.
化学聚集电融合法在新型酵母构建中的应用   总被引:1,自引:1,他引:1  
在细胞融俣过程中化学聚集电融合法,即用PEG作为聚合剂与电融合法连用,取得明显的效果。其条件是:电压11kV;脉冲宽度5μ脉冲个数3;脉冲时间59s;温度4℃,PEG浓度35%;自理时间30min;处理温度30℃及pH6.从细胞体系、DNA含量,淀粉糖化能力及区区糖发酵能力等方面说明,所获得的新型酵母是两亲本的融合株。  相似文献   

4.
用促黄体激素(LH)为抗原免疫BALB/C小鼠,通过免疫小鼠脾细胞和SP_2/0小鼠骨髓瘤细胞的融合,产生分泌抗LH单克隆抗体(McAbs)的杂交瘤细胞。用ELISA法检测融合细胞培养上清,从195份检样中测出两份能与LH抗原起反应。对该2孔杂交瘤细胞进行亚克隆和扩大培养。经体外四个月传代培养及液氮冻存后复苏培养。仍保持分泌抗LH的McAbs的能力,所建立的二株杂交瘤细胞命名为AL—01和AL—02。给予注液体石蜡的BALB/C小鼠注入该二种细胞,均能诱生腹水。用ELISA法测腹水抗体,效价分别为10~(-6)以上和10~(-5)以上。通过染色体组型分析,确认AL—01和AL—02皆为杂交瘤细胞。  相似文献   

5.
采用重组小鼠凝血因子Ⅸ制备单克隆抗体的研究   总被引:2,自引:0,他引:2  
将小鼠凝血因子Ⅸ (mFⅨ )cDNA蛋白编码区序列克隆至原核表达载体 pQE30 ,在大肠杆菌M 15中获得高效表达 (约占细菌蛋白总量的 5 1 2 % ) ,并经过SDS PAGE纯化获得均一的重组mFⅨ蛋白 .以此蛋白为抗原 ,免疫Lou/MN系大鼠 ,取其脾细胞与小鼠骨髓瘤细胞 (Sp 2 / 0 )融合后 ,经 3次克隆化选择获 2株杂交瘤细胞(H4B5和C2B10 ) .以 1× 10 6杂交瘤细胞注射免疫缺陷型裸鼠 ,10d后获得腹水 .对此单抗的功能和特异性初步研究表明 ,所制备的抗小鼠FⅨ单抗可用于Western免疫印迹等研究 .  相似文献   

6.
为了对伊氏锥虫病的免疫诊断提供有价值的单克隆抗体,笔者通过了3次SP2/0骨髓瘤细胞与伊氏锥虫抗原免疫的BALB/c小鼠脾细胞间的融合试验和多次筛选以及克隆化,建立起7株分泌单克隆抗体的杂交瘤细胞株。对其中ⅡG_6杂交瘤作了进一步的检定分析,用ⅡG_6上清液对T.e.抗原进行ELISA试验,显示出高度的特异性,与血吸虫、弓形虫抗原均未出现交叉反应。ELISA测试ⅡG_6细胞株培养上清效价为1:5.1×10~3,而杂交瘤细胞接种BALB/c小鼠诱生腹水抗体的效价为1:1.6×10~7。该株单克隆抗体免疫球蛋白的类型鉴定属IgG_1亚类。该株细胞已在体外培养下,稳定地分泌特异性抗体至少达6个月之久,同时在液氮冻存近8个月之后仍能保持分泌抗体的能力。  相似文献   

7.
体细胞电融合技术因其融合效率高、对细胞无毒害作用而愈来愈普遍地用于细胞工程技术操作中。为了准确地确定实验参数,试验采用一个组装的电融合仪和自制平行电极,以马铃薯和薯蓣为材料,研究了在不同原生质体密度和不同电场强度条件下的细胞融合效率。试验结果显示,以交变电场频率1.0MHz、电压140Vpp.cm-处理原生质体20-40秒,尔后施加瞬时直流高压(120kv)脉冲1-2次,可以获得73.5%─76.7%的融合细胞,其中11.5%─14.4%的融合细胞可发生正常分裂。在本试验设计范围内,电场强度对细胞融合频率的影响主要表现在融合细胞能否发生分裂,而初始融合频率在各处理间无显著差异。  相似文献   

8.
将小鼠骨髓瘤细胞(SP2/0细胞)与经TCS免疫过的小鼠脾细胞融合,融合后的细胞悬浮于HAT培养液中进行选择性培养,挑选可以分泌TCS单克隆抗体的细胞系,再经克隆化为单克隆细胞系。用间接酶联免疫吸附法检测此细胞系分泌单克隆抗体情况。经细胞融合和2次克隆化后,获得1株可以稳定分泌TCS单克隆抗体杂交瘤细胞系,ELISA检测结果为阳性。获得了可以稳定分泌天花粉蛋白单克隆抗体的杂交瘤细胞系,为TCS鉴定、应用奠定良好的基础。  相似文献   

9.
抗爪蟾角蛋白单克隆抗体的研究   总被引:1,自引:1,他引:0  
本实验以非洲爪蟾(Xenopus laevis)成体皮肤角蛋白免疫的BALB/c鼠脾细胞与小鼠Sp2/0骨髓瘤细胞在PEG的作用下进行融合,经HAT选择培养、ELISA筛选、反复克隆化及冻存复苏,获得四株稳定分泌抗体的杂交瘤细胞株。对其中一株杂交瘤细胞KD_(10)及其分泌的抗体进行了较全面的研究。  相似文献   

10.
介绍了相控线阵声波测井辐射器的工作原理,根据辐射器各个阵元的性能参数设计了两种供实验使用的激励信号源,即5通道Burst信号源和4通道脉冲信号源。Burst信号源输出电压峰值连续可调(6~30 V),相邻通道之间的延迟时间可调(0~22μs),步长为1μs,输出频率可根据换能器的频带在10~250 kHz选择。脉冲信号源输出电压幅度连续可调(50~150 V),延迟时间为0~7μs可调,步长为0.25μs,脉冲宽度为1~4μs。利用这两种信号源可以对相控线阵声波辐射器进行相位和幅度加权控制。在实验室内对这两种信号源进行了性能测试,结果表明,两个信号源均能满足实验要求。  相似文献   

11.
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.  相似文献   

12.
AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…  相似文献   

13.
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。  相似文献   

14.
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  相似文献   

15.
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  相似文献   

16.
17.
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.  相似文献   

18.
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).  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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