成丝的速度与方式和蜘蛛丝力学性能的关系

为了分析蜘蛛丝的力学性能和成丝条件间的关系,研究了不同速度下人工卷取的蜘蛛牵引丝以及蜘蛛垂直下落时分泌的牵引丝的力学性能.研究结果表明,随着卷取速度的增大,蜘蛛丝的断裂强度有一极大值,在高速卷取时强度的变化较低速时平缓.垂直下落时蜘蛛分泌的牵引丝的综合力学性能优异,但断裂强度不一定大于人工卷取的丝.成丝速度和成丝方式都对蜘蛛丝的性能有很大影响.     

蜘蛛丝的结晶结构及其取向

研究和分析了大腹园蛛的牵引丝、内层包卵丝的结晶结构及其取向,探索了大腹园蛛丝纤维分子链的排列状态、结晶结构和蜘蛛丝力学性能间的关系。     

大腹圆蛛包卵丝的化学组成与物理机械性能

分析了大腹圆蛛包卵丝的氨基酸组成，包卵丝中极性氨基酸含量较高，小侧基氨基酸含量较少，利用SEM及计算机图像处理技术分析了大腹圆蛛包卵丝的形态结构特征，包卵丝的断面形状基本为圆形，包层包卵丝的细度约为内层包卵丝的2倍，并且外层包卵丝的纤维断面内有许多无规分布的纳米级孔隙，经冷冻干燥后，孔隙率增加了2倍左右，在此基础上，研究了大腹圆蛛包卵丝的拉伸机械性能，光泽和热性能，初步探索了性能的形成机理，大腹圆蛛内层包卵丝具有比其他功能蜘蛛丝和丝素纤维好得多的力学性能，其强度高，伸长率大，韧性好，包卵丝在光泽和热性能方面与蚕丝丝素纤维有一定的差异。     

大腹圆蛛拖丝蛋白基因3'端克隆和序列分析

用PCR(Polymerase Chain Reaction)技术从大腹圆蛛主壶腹腺组织的基因组DNA和RNA中分别扩增获得编码拖丝蛋白基因的3'端基因序列,克隆PCR产物到pDrive载体,经PCR、限制性酶切筛选和序列分析后证明获得了蜘蛛拖丝蛋白基因两个克隆AvDS1和AvRS2.基因序列和氨基酸序列分析表明AvDS1和AvRS2具有拖丝蛋白基因序列和蛋白质氨基酸序列特有的结构特征,氨基酸序列中存在4个motifs-(A)n(GA)n(GFGX)n以及(GGX)n.AvDS1与基因库中已报道的蜘蛛拖丝蛋白基因序列同源性较低,而AvRS2与已经报道的Nephila clavipes拖丝蛋白基因组分-1的同源性可达91%,推测AvRS2是编码大腹圆蛛拖丝蛋白基因组分-1基因片段.比较这两个克隆的核苷酸序列和氨基酸序列,发现它们之间没有同源性.推测它们是同一基因的不同片段,或是编码大腹圆珠拖丝蛋白的两个基因组分. 明AvDS1和AvRS2具有拖丝蛋白基因序列和蛋白质氨基酸序列特有的结构特征,氨基酸序列中存在4个motifs-(A)n(GA)n(GFGX)n以及(GGX)n.AvDS1与基因库中已报道的蜘蛛拖丝蛋 基因序列同源性较低,而AvRS2与已经报道的Nephila clavipes拖丝蛋白基因组分-1的同源性可达91%,推测AvRS2是编码大腹圆蛛拖丝蛋白基因组分-1基因片段.比较这两个克隆的核苷酸序列和氨基酸序列,发现它们之间没有同源性.推测它们是同一基因的不同片段,或是编码大腹圆珠拖丝蛋白的两个基因组分. 明AvD     

重组拖丝蛋白基因在大肠杆菌中表达条件的优化

对巳构建的重组蜘蛛拖丝蛋白基因表达菌株pNS2，以IPTG为诱导剂，建立最适表达条件．在LB培养基中添加质量分数为0．1％的甘氨酸和丙氨酸，菌体培养至密度A600＝0.5—0．7时加入浓度为0．2mmol/L的IPTG，诱导5h，可得到表达量占可溶性总蛋白质27．2％的较好结果．     

蜘蛛丝蛋白的研究进展

蜘蛛丝属线状蛋白质,具有独特的结构和性能特点,是世界上拉力强度和弹性最强的纤维之一,其强度比钢还要硬.一些蜘蛛丝蛋白的基因已被克隆出来,并且用不同的表达载体表达,同时进行了成丝研究.由于蜘蛛丝性能独特,在许多领域具有良好的应用前景.     

蜘蛛丝多肽链段结构的优化设计

根据蜘蛛丝氨基酸片段的序列特征,利用 Hyperchem5软件对部分具有β-折叠或螺旋状结构的多肽链段的分子结构进行优化设计,分析了氨基酸种类、聚合度对多肽链段形成特定构象的影响,发现一定的氨基酸片段有形成特定构象的趋势。分子构象的多元化是蜘蛛丝具有优异力学性能的重要原因。     

Conformational Transitions and Self-assembly of Regenerated Spider Silk Protein in Water

Spider silk, relying on its exceptional mechanical properties, has attracted extensive attention throughout the world. The structure of a material can influence its mechanical properties. Investigation of the structure of spider silk includes amino acid composition, molecular structure, self-assembly, and crystallization, among other characteristics. Herein, the effects of concentration, time, alkali metal ions （Na^＋and K ^＋ ） and pH on the conformational transition and self-assembly of regenerated Ornithoctonus huwena spider dragline silk protein （spidroin） in water were investigated using circular dichroism （CD） and atomic force microScopy （AFM）. Spidroin concentration, time and Na ＋ ions slightly influenced the conformational transition of spidroin molecules. However, K ＋ ions and pH induced the formation of a β- sheet structure. Increasing spidroin concentration or time increased the aggregation of spidroin and enhanced the formation of nanoffiaments. K ~ ions enhanced the serf-assembly of spidroin into nanofilaments. The self-assembled nanofilaments appeared at a pH of approximately 6. 11. Both lower and higher pH induced aggregation. At a lower pH, the aggregation was composed of nanopartides, whereas higher pH induced the aggregation of nanoffiaments, likely from the synergistic effect of Na ^＋ ions and pH.     

蜘蛛丝拉伸变形行为的力学模型

根据大腹园蛛牵引丝力学性能的结构机理及其聚集态结构和形态结构特征，初步建立了以皮芯层结构为基础的蜘蛛丝拉伸力学模型，分析了皮芯层比例及结构对蜘蛛丝纤维力学性能的影响。以层状复合材料的拉伸变形为依据，分析了蜘蛛丝纤维的拉伸断裂过程与皮芯层性能间的关系。     

Molecular Fundaments of Mechanical Properties of Spider Silk

Dragline, framework and cocoon silk fibers of Araneus Ventricosus were used for this study. To investigate the microstructure mechanisms of stress-strain behavior of spider silk, firstly, amino acid compositions were analyzed and molecular conformations and crystallinity were measured with Raman spectra and X-ray diffraction respectively. The results showed that there were more amino acids with large side groups and polar ones in spider silk than those of Bombyx silk, and the amino acid distribution varied with different spider silk. The molecular structures were mainlyα-helix and β-sheet, and random coil andβ-turn existed as well. The proportions and arrangement of these conformations of dragline silk were different from framework and cocoon silk fibers. Microstructure was one of important factors of excellent mechanical properties of spider silk. Crystallinity of spider silk was very low, which implied that the roles of crystal on spider silk were not as great as other protein fibers.     

蜘蛛丝应力—应变行为的分子结构机理

利用激光拉曼光谱技术研究了大腹园蛛牵引丝、蛛网框丝和内层包卵丝的分子构象,分析了蜘蛛丝的分子构象差异对其应力-应变行为的影响,探索了蜘蛛丝优异力学性能的分子结构机理。     

基因重组蛛丝蛋白仿生纤维的制备研究

以甲酸为溶剂,制备质量浓度为70 g/mL基因重组蛛丝蛋白纺丝液.分别以甲醇、硫酸铵为凝固浴,经一段拉伸、二段拉伸制备仿生纤维,分析仿生纤维的水溶性、密度、比强度等物理和机械特性,并考察加入高分子材料聚乙烯醇及纤维后处理对纤维性能结构的影响.     

大腹圆蛛毒素的双向电泳分析

提取大腹圆蛛毒素,进行双向凝胶电泳(Two Dimension Gel Electrophoresis,2-DGE),经银染法显色及电脑软件分析,结果表明:大腹圆蛛的毒素至少有80种成份,其中大部分显碱性或中性.     

早稻田蜘蛛群落结构及优势种分析

通过稻田定点调查,初步鉴定长沙地区稻田有蜘蛛8科28种,其中主要成分依次为狼蛛、皿蛛、球蛛和肖蛸等;蜘蛛群落结构在水稻的不同发育期有明显不同;根据多样性指数（H）、优势度（C）和均匀度（E）的分析,早稻发育前期（5月）蜘蛛群落物种多样性明显小于中期（6月）和后期（7月）。优势种为拟水狼蛛和食虫沟偏蛛,分析了拟水狼蛛和食虫沟瘤蛛及蜘蛛混合种群的消长动态,对优势种蜘蛛与目标害虫的关系进行了初步分析。     

Local conformation transitions of linear DNA induced by cisplatin

Cisplatin is the most successful anti-tumor drug, and its pharmacological property is generally considered to derive from the modification of DNA molecules. Structural modifications of short DNA induced by cisplatin have already been investigated. However, the conformation transitions induced by cisplatin are not clear. In the present letter, we have studied the effect of low-concentration cisplatin on DNA conformation by using AFM imaging. We observed formations of micro-rod structures of linear DNA induced by cisplatin. A method is presented to quantitatively analyze the occurrence of micro-rod structures. We found that the formation of micro-rod structures depends on the DNA sequence. Based on the results, we proposed a physical mechanism to explain the local conformation transitions of DNA molecules under the influence of cisplatin.     

RGD-拖丝蛋白基因的构建及其在毕赤酵母中的分泌表达

根据天然拖丝蛋白的高度重复性序列,引入与细胞黏附有关的精氨酸-甘氨酸-天冬氨酸(RGD)三肽序列,以毕赤酵母偏好的密码子化学合成RGD-拖丝蛋白基因单体,通过"头尾相连"的多聚化策略,倍加成16聚体与32聚体基因.分别将这两种多聚体与分泌型表达载体pPIC9K连接,转化毕赤酵母GS115,用G418筛选毕赤酵母重组菌.通过甲醇诱导表达,培养液上清的SDS-PAGE分析表明RGD-重组拖丝蛋白获得分泌型表达.     

蚕丝的开发再利用

就国内外蚕丝的研究与应用现状进行综述，着重概括近年来蚕丝在 只、医药生物、食品、化妆品等领域的开发再利用情况，指出蚕丝在非衣料领域的广阔应用前景。     

HPLC-MS法分析玉米须中黄酮苷类化合物

研究了玉米须中的黄酮苷类成分。通过硅胶柱层析和Sephadex LH-20层析对玉米须乙酸乙酯提取物初步纯化,采用液相色谱-质谱联用法对其中的黄酮苷类化合物进行分析。结果表明,玉米须乙酸乙酯提取物中含有3种黄酮苷类化合物,分别为5,7,4′-三羟基-3′-甲氧基黄酮-6-C-2″-O-二鼠李糖苷、5,7,4′-三羟基-3′-甲氧基黄酮-3-C-阿拉伯糖-6-C-鼠李糖苷和ax-5″-甲烷-3′-甲氧基玉米素。     

Progress in measuring biophysical properties of membrane proteins with AFM single-molecule force spectroscopy

Membrane proteins are crucial in cell physiological activities and are the targets for most drugs.Thus,investigating the behaviors of membrane proteins not only provide deeper insights into cell function,but also help disease treatment and drug development.Atomic force microscopy is a unique tool for investigating the structure of membrane proteins.It can both image the morphology of single native membrane proteins with high resolution and,via single-molecule force spectroscopy(SMFS),directly measure their biophysical properties during molecular physiological activities such as ligand binding and protein unfolding.In the context of molecular biomechanics,SMFS has been successfully used to understand the structure and function of membrane proteins,complementing the static three-dimensional structures of proteins obtained by X-ray crystallography.Here,based on the authors’antigen-antibody binding force measurements in clinical tumor cells,the principle and method of SMFS is discussed,the progress in using SMFS to characterize membrane proteins is summarized,and challenges for SMFS are presented.     

有机溶剂对纳滤膜结构及透过特性的影响

观察了两种纳滤膜(NF-SH,MPF-44)的稳定性,并利用原子力电镜技术(AFM)观察膜的活性表面结构,发现有机溶剂导致膜结构发生显变化。观测在溶剂中的传递通量以及溶质截留率发现,影响NF-SH膜的通量的主要因素是膜本身的结构;对MPF-44膜,影响膜通量的主要因素是溶剂性质。对于这两种纳滤膜影响截留率的主要因素都是溶剂与溶质的相互作用。