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171.
双水相电泳分离蛋白质的研究 总被引:2,自引:0,他引:2
近几年来,随着生物技术的迅速发展,制备型电泳技术的研究得到了重视。然而由于技术上的原因,大规模的制备型电泳技术的研究还未能取得突破。阻碍电泳放大的一个主要问题是由于电加热作用而导致的热对流对电泳分离的破坏。为解决这一问题,人们提出了许多方法。例如,在太空的微重力环境下进行电泳,应力稳定自由流动电泳,循环等电聚焦和区带电泳,色谱电泳和等电膜等电聚焦等。这些方法在电泳放大上都取得了一定的进展,但各有其局限性。最近,Clark提出利用双水相的液液界面阻止热对流的设想,为开发大规模的制备型电泳技术开辟了一条新途径、Raghava Rao等在两种双水相体系上施加电场后成倍地缩短了分相时间。Levine和Bier采用U型管电泳装置研究了双水相体系中血红蛋白的电泳迁移率,观测到界面有阻滞作用。Clark在柱型电泳装置中进行了一组双水相萃取肌红蛋白的简单实验。在10mA的恒电流下电泳40min之后,肌红蛋白的分配系数为7.5,而当电场反向后,分配系数变为0.04,界面阻力并不显著,两者结论并不一致。 相似文献
172.
173.
用原子力显微镜(AFM)研究了磷脂DMPC三层Langmuir-Blodgett(LB)膜的分子排列结构,结果表明:在磷脂LB膜的两相(液体压缩相Liquid-condensedphase和液体扩张相Liquid-expandedphase)共存时,液体压缩相中的磷脂分子排列紧密,取向一致,分子间作用力较大,因而能够得到分子图像。而液体扩张相中的磷脂分子排列松散,取向混乱。分子间的作用力较弱,难于得到分子图像。在液体压缩相中磷脂分子以单斜晶格结构排列,分子间隔为0.72nm.分子高度为2.1nm。这一结果和DMPC的单晶结构进行了比较。 相似文献
174.
175.
鹅掌楸雌配子体败育对生殖的影响 总被引:12,自引:0,他引:12
胚珠和雌配子体败育是限制鹅掌楸生殖成功的一个重要因素。中国东部和西部鹅掌楸种群在雌配子体发育的各阶段上的败育程度有差异,以西部种群的发育较好。西部分布区较合适的生境促进了胚囊的发育,一定温度和湿度的环境可以活化珠心细胞输送营养物质供给雌配子体发育,提高受精和结籽的能力 相似文献
176.
用自制的氨基PEG化试剂rIL-2进行化学修饰,研究了试剂浓度,溶液pH,反应时间等与PEca-rIL-2产率及IL-2活性保持之间的关系,建立了一套获得稳定修饰度的PEG-rIL-2的方法。研究发现,反应时间跟修饰度关系不大;溶液pH对修饰度有一定的影响,中性pH以上反应都可进行;而试剂浓度直接决定修饰度的高低,过量越多,修饰度越高,而生物活性保留也越低;但低度修饰,对活性几乎没有影响,可保留活性在95%左右。 相似文献
177.
Conservation of an intact human immunodeficiency virus type 1 vif gene in vitro and in vivo. 总被引:5,自引:5,他引:0 下载免费PDF全文
P Sova M van Ranst P Gupta R Balachandran W Chao S Itescu G McKinley D J Volsky 《Journal of virology》1995,69(4):2557-2564
Replication of vif-negative human immunodeficiency virus type 1 (HIV-1) is attenuated in certain cell lines and highly impaired in peripheral blood lymphocytes in vitro. To determine whether intact vif is positively selected during natural HIV-1 infection and to determine vif sequence variability, we employed PCR amplification, cloning, and sequencing to investigate the vif region of replicating virus in short-term-passage HIV-1 primary isolates from five asymptomatic individuals and from five persons with AIDS. A total of 46 vif clones were obtained and analyzed. Recombinant proviruses were constructed from selected vif clones from one patient and found to be fully infectious. We found that 38 of the 46 clones sequenced carried open vif reading frames and that there was a low degree of heterogeneity of vif genes within isolates from the same individual and among isolates from different donors. The cysteines previously found to be essential for vif protein function were conserved in all clones. A phylogenetic tree constructed from all available vif nucleotide sequences resulted in a virus grouping similar to those of gag and env. Direct sequencing of vif amplified by PCR from uncultured lymphocytes of 15 individuals at various stages of progression toward AIDS demonstrated vif open reading frames in 13 of 15 samples tested. There was no obvious correlation between disease status and the presence of an intact vif within this sample group at the time of sample procurement. The conservation of the vif open reading frame in vitro and in vivo and its limited variability following virus transmission in vitro are consistent with a role for vif in natural HIV-1 infection. 相似文献
178.
Aberrant Gag protein composition of a human immunodeficiency virus type 1 vif mutant produced in primary lymphocytes. 总被引:7,自引:6,他引:1 下载免费PDF全文
Productive, spreading infection of peripheral blood lymphocytes (PBL) with human immunodeficiency virus type 1 (HIV-1) requires the viral protein Vif. To study the requirement for vif in this system, we infected PBL with a phenotypically complemented HIV-1 clone mutated in vif. Progeny virus was produced which was noninfectious in PBL but replicated in SupT1 cells. Analysis of metabolically labeled proteins of sedimentable extracellular particles made in PBL by radioimmunoprecipitation with either serum from a patient with AIDS or a monoclonal antibody reactive with HIV-1 Gag proteins revealed that vif-negative but not wild-type particles carry higher levels of p55, p41, and p38 Gag-specific proteins compared with those of p24. Similar results were obtained with sucrose-purified virions. Our data indicate that vif plays a role in Gag protein processing or in incorporation of processed Gag products into mature virions. The presence of unprocessed precursor Gag polyprotein (Pr55gag) and other Gag processing intermediates in PBL-derived vif-negative extracellular particles may contribute to the reduced infectivity of this virus. 相似文献
179.
The Impact of Chlorophyll-Retention Mutations,d1d2 and cyt-G1, during Embryogeny in Soybean 下载免费PDF全文
The ultrastructural, physiological, and molecular changes in developing and mature seeds were monitored in a control line (Glycine max [L.] Merr., cv Clark) that exhibited seed degreening and two mutant lines (d1d2 and cyt-G1) that retained chlorophyll upon seed maturation. Ultrastructural studies showed that the control line had no internal membranes, whereas stacked thylakoid membranes were detected in the green seed from the mutant lines. Pigment analyses indicated that total chlorophyll was lowest in the mature seeds of the control line. Mature d1d2 and cyt-G1 seed had elevated Chl a and Chl b levels, respectively. In both control and mutant lines, Lhcb1, Lhcb2, and RbcS mRNAs were abundant in embryos prior to cotyledon filling, declined after the onset of storage protein accumulation, and were barely detectable or undetectable in all later stages of seed development. Therefore, the chlorophyll-retention phenotype must be a result of the alteration of a process that occurs after translation of photosynthesis-related mRNAs to stabilize apoprotein and pigment levels. Furthermore, different elements controlling either the synthesis or turnover of Chl a and Chl b must be impaired in the d1d2 and cyt-G1 lines. No reproducible differences in total leaf, embryonic, and chloroplast protein profiles and plastid DNAs could be correlated with the mutations that induced chlorophyll retention. 相似文献
180.