淀粉凝胶制备电泳

电泳是分离蛋白质等物质的有效方法之一,电泳分析己经普遍应用,但是,由于电泳时热效应的产生和浓度梯度的产生,使电泳用于制备发生了困难。以凝胶为支持介质的电泳可减少连续液流电泳所存在的对流问题,再采取适当冷却措施就可进行制备电泳。     

一种分析有丝分裂期蛋白的新方法

介绍了一种分析细胞有丝分裂任一时期蛋白质组成及各时期蛋白动态变化的方法，该法将细胞的显微分离，微量蛋白电泳和银染结合起来，通过显微分离可以获得无期它时期细胞污染的任一时期细胞，通过微量蛋白电泳和银染，可以分析其蛋白组成，并通过显微分离100个小麦根尖末期细胞证明了其可行性。     

蛋白质印迹技术在生物医学中的应用方法

蛋白质印迹（Westem blot）技术是通过聚丙烯酰胺电泳根据分子量大小分离蛋白后转移到杂交膜上，然后通过一抗／二抗复合物对靶蛋白进行特异性检测的方法。蛋白质印迹技术进行蛋白质分析最流行和作成熟的技术之一，超过60％的生命科学工作者使用该方法。     

用泡沫分离法浓缩和分离蛋白质(Ⅰ)——设备及分离过程的研究

提出一种新型泡沫分离设备即环流泡沫塔，以牛血清清蛋白（ＢＳＡ）的分离过程为例，对比了在环流泡沫塔与鼓泡塔中泡沫分离蛋白质的动力学行为，考察了操作参数（气速）、溶液体系的性质（ｐＨ和离子强度）对分离过程的影响。以表面张力作为描述蛋白质在水溶液中的表面活性的参量，研究了蛋白质溶液体系性质对其表面张力和泡沫分离效果的影响。结果表明，在接近蛋白质等电点处，溶液的表面张力最低，进行泡沫分离效果最佳。     

蛋白质分离纯化与层析技术进展

蛋白质分离纯化是蛋白质产品工业化生产的关键之一。文中分析了蛋白质分离纯化的特点，指出了分离纯化蛋白质应解决的一些问题。层析技术是一种有效的蛋白质纯化方法，文中从层析方法、层析固定相、层析过程的数学模拟三方面评述了层析技术的新进展     

亲和温敏聚合物PNIPAA,-co-AA,的制备及表征

制备一种新型的蛋白质分离纯化材料。探讨亲和温敏双水相体系对牛血清白蛋白（BSA）的分配特性。优化分离纯化BSA条件,同时测定BSA的回收率以及纯化倍数,并通过电泳SDS-PAGE对BSA蛋白进行了表征。     

焦耳热对芯片电泳分离效率影响的模拟分析和实验

采用CoventorWare有限元分析软件对电泳芯片管道内焦耳热效应进行了模拟计算和分析,以牛血清白蛋白和溶菌酶作为样品分析对象,通过考虑芯片不同绝缘层厚度、分离场强、缓冲液浓度条件下分离效能,来考察芯片电泳过程中管内焦耳热现象对样品分离的影响,明确了在综合电泳分离条件中限制焦耳热的必要性。     

橡胶草叶片蛋白质提取方法的选择及双向电泳体系的优化

目的为了建立起橡胶草叶片组织的蛋白双向电泳体系。方法采用5种不同方法提取橡胶草叶片蛋白,并通过蛋白提取率和单向SDS-PAGE电泳的比较,筛选出改良TCA-丙酮沉淀法、酚-乙酸铵沉淀法、Tris浸提法3种方法进行2-DE电泳。结果表明选择改良TCA-丙酮法为最合适的蛋白质提取方法。为进一步优化体系,比较了不同染色方法、上样量和固相p H梯度预制胶条(IPG)的p H范围对双向电泳结果的影响,电泳结果表明使用17 cm p H 3-10的IPG胶条、上样量为1200μg、考马斯亮蓝染色法能分离得到更多的蛋白点,是橡胶草叶片双向电泳体系的最优条件。结论本研究初步建立了橡胶草叶片蛋白质双向电泳体系,为深入研究橡胶草蛋白质组学提供依据。     

种子蛋白电泳技术

种子蛋白电泳是指通过特定的电泳方法对种子蛋白进行分离和鉴别的一种技术,最常用的是酸性聚丙烯酰胺凝胶电泳(A-PAGE)和SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)。聚丙烯酰胺凝胶电泳作为一种高分辨率的分析鉴定技术,在蛋白质、多肽、核酸等生物大分子的分析中一直广泛使用,通过种子蛋白电泳获得的指纹图谱在测定良种质量(真伪、纯度)防止伪劣种子流入市场,保护我国名、优、特种质资源及育成的知识产权和育种家们的权益等方面,均具有重要意义。     

中国柞蚕血淋巴的盘状电泳

应用聚丙烯酰胺凝胶盘状电泳法对中国柞蚕血淋巴中的蛋白质进行分离．实验结果表明，昆虫血淋巴中的蛋白质种类远较高等生物和人的少，而且还发现不同品种间的柞蚕血淋巴中蛋白质种类和分子大小也不同，从而为蚕的分类学和饲养提供了理论根据     

Molecular heterogeneity of benzodiazepine receptors

Benzodiazepines exhibit reversible, stereospecific high affinity binding to mammalian brain membranes, and the respective binding sites for 3H-flunitrazepam represent pharmacologically and clinically relevant receptors for benzodiazepines. Recently it has been demonstrated that reversibly bound 3H-flunitrazepam becomes irreversibly attached to a specific membrane protein with apparent molecular weight of 50,000 when incubations are performed in the presence of UV light. Irreversible binding of 3H-flunitrazepam to this protein had pharmacological properties similar to reversible benzodiazepine receptor binding, indicating that 3H-flunitrazepam is a photoaffinity label for the benzodiazepine receptor. Using irreversible binding of 3H-flunitrazepam and subsequent electrophoretic separation of the labelled proteins in SDS-gels followed by fluorography, we found that in hippocampus and several other brain regions at least two different types of benzodiazepine receptors exist. Each seems to be associated with a gamma-aminobutyric acid (GABA) receptor.     

Multiple conformations of a protein demonstrated by magnetization transfer NMR spectroscopy

It is generally accepted that a globular protein in its native state adopts a single, well-defined conformation. However, there have been several reports that some proteins may exist in more than one distinct folded form in equilibrium. In the case of staphylococcal nuclease, evidence for multiple conformations has come from electrophoretic and NMR studies, although there has been some controversy as to whether these are actually interconvertible forms of the same molecular species. Recently, magnetization transfer (MT)-NMR has been developed as a means of studying the kinetics of conformational transitions in proteins. In the study reported here, this approach has been extended and used to demonstrate the presence of at least two native forms of nuclease in equilibrium and to study their interconversion with the unfolded state under the conditions of the thermal unfolding transition. The experiments reveal that two distinct native forms of the protein fold and unfold independently and that these can interconvert directly as well as via the unfolded state. The spectra of the different forms suggest that they are structurally similar but the MT experiments show that the kinetics of folding and unfolding are quite different. Characterization of this behaviour will, therefore, have important implications for our understanding of the relationship between structure and folding kinetics.     

水力机械三维分离准则的探讨

水力机械的分离为三维流动，三维流动的侧向压力梯度产生边界层的二次流动。文中将边界层内的速度分布分为近壁粘性层和接近于主流的外层两部分，通过研究垂直于主流方向的二次流动和主流的相关性，结合三维分离线的邻近流动特性分析提出了通过主流流场参数和沿流向的二维边界层流动来判断三维流动分离的准则。在垂直于三维分离线的截面内，其流动性态和二维分离相类似。边界层的二次流动和侧向压力梯度，以及二者的相互作用是影响水力机械三维流动分离的重要因素。当主流逆压梯度的方向与二次流动的方向均在主流方向一侧时，流动的三维效应使得三维流动比二维流动不容易分离。当主流逆压梯度的方向与二次流动的方向分别在主流方向的两侧时，三维流动较易产生分离     

假丝酵母属可溶性蛋白及酯酶的聚丙烯酰胺凝胶电泳

本文对分离自黄海海水的无名假丝酵母Ｃａｎｄｉｄａｆａｍａｔａ（７株）、季也蒙假丝酵母Ｃａｎｄｉｄａｇｕｉｌｌｉｃｌｍｏｎｄｉｉ（５株）、近平滑假丝酵母Ｃａｎｄｉｄａｐａｒａｐｓｉｌｏｓｉｓ（５株）进行了可溶性蛋白及酯酶的聚丙烯酰胺凝胶电泳。经多次实验，三个种均恒定地呈现典型的蛋白及酯酶图谱。不同种间的谱带差异较大，同种不同株间谱带相似。     

Multiple forms of dynamin are encoded by shibire, a Drosophila gene involved in endocytosis.

Dynamin was discovered in bovine brain tissue as a nucleotide-sensitive microtubule-binding protein of relative molecular mass 100,000. It was found to cross-link microtubules into highly ordered bundles, and appeared to have a role in intermicrotubule sliding in vitro. Cloning and sequencing of rat brain dynamin complementary DNA identified an N-terminal region of about 300 amino acids which contained the three consensus elements characteristic of GTP-binding proteins. Extensive homology was found between this domain and the mammalian Mx proteins which are involved in interferon-induced viral resistance, and with the product of the VPS1 locus in Saccharomyces cerevisiae, which has been implicated both in membrane protein sorting, and in meiotic spindle pole separation. Dynamin-containing microtubule bundles were not observed in an immunofluorescence study of cultured mammalian cells, but a role for a GTP-requiring protein in intermicrotubule sliding during mitosis in plants has been reported. We report here that Drosophila melanogaster contains multiple tissue-specific and developmentally-regulated forms of dynamin, which are products of the shibire locus previously implicated in endocytic protein sorting.     

Recent Advances in Protein Extraction and Chiral Separation of Blomolecules

Reverse micelles create unique environment in organic media. They are capable of solubilizing hydrophilic biomolecules （e.g., proteins, peptides, amino acids, and DNAs） in their aqueous interior. This feature brings about the practical use of biomaterials in organic media because reverse micelles solubilize them with the intrinsic activity. In this paper, we focus on recent two topics concerning protein extraction and chiral separation of biomolecules using liquid membranes. In the first topic, we present recent attempts to extract proteins from an aqueous solution into isooctane using reverse micelles, and some important operational parameters to achieve an efficient protein transfer are discussed. Furthermore, novel function of reverse micelles as a protein activation medium is introduced. In the reverse micellar phase, denatured proteins were completely reactivated in the reverse micellar solution. The reverse micellar technique is found to be a useful tool not only for protein separation but also for protein refolding. Furthermore, we found that a cyclic ligand carixarene has an extraction ability to set up optimum conditions for protein transfer. In the second topic, we have found that a supported liquid membrane （SLM） encapsulating enzymes shows high enantioselectivity （enantioselective excess value is over 96%） in the transport of racemic pharmaceutical compound ibuprofen. A different experiment also suggests that the α-chymotrypsin-catalyzed reactions droved the enantioselective transport of L-phenylalanine based on the enantioselectivity of the enzyme. The SLM encapsulating the surfactant-enzyme complex enabled the highly enantioselective separation of racemic mixtures. It can be envisioned that arrangement of appropriate enzymes in the SLM system will allow enantioselective separation of various useful organic compounds.     

边界层分离点实时数据采集与处理系统的设计与实现

通过分析实时数据采集与处理的基本原理和系统构架,搭建了适用于微型飞行器边界层分离点检测的实时信号采集与处理系统.该系统主要由模拟低通滤波电路、模数转换电路(ADC)、场可编程门阵列(FPGA)控制电路及USB接口电路构成.测试结果为:基准电平0.1mV,峰峰值407mV,频率1kHz,满足边界层分离点实时检测和判定的要求.     

多功能蛋白增活器的研制及性能研究

自制了一种新型的多功能蛋白增活器,它具有同时除变性剂、分离杂蛋白、复性目标蛋白及便于回收变性剂的功能。考查了其对蛋白的分离、复性性能。发现这种多功能蛋白增活器可对６种标准蛋白进行良好地分离。同时对盐酸胍变性的溶菌酶和核糖核酸酶完全复性。通过测定压力－流速曲线,发现多功能蛋白增活器的压力均远远低于普通色谱柱。将其应用于基因工程发酵的粒细胞集落因子（Ｇ－ＣＳＦ）的分离纯化,可使纯度接近于１００％,生物活性大于常规方法３倍     

聚丙烯酰胺凝胶电泳——曙红Y染色法

十二烷基硫酸钠—聚丙烯酰胺凝胶电泳已广泛用于分子生物学和医学临床等领域,但该技术的染色方法多沿用考马斯亮蓝和银染色,这两种方法均有其局限性,如蛋白凝胶染色前须经酸或醛固定,不易洗脱且操作繁琐,脱色耗时等。而曙红Y染色具有检测灵敏度高,操作简便,染色时间短,可对各种蛋白质染色,染色后的蛋白质可直接进行电泳转移印迹,也可将凝胶中蛋白质电泳转移后染色。染色后的蛋白质仍具有抗原性。此外,还具有试剂便宜,使用安全,便于推广应用等优点。