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     

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.     

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。     

Morphology and Microstructure of Spider Dragline Silk from Araneus Ventricosus

The spider dragline silk has excellent mechanical properties. The stress- strain curves of dragline silk fibers have intraspecific and intraindividual variability because of the spiders active control during spinning process. To investigate the relationship between the morphology of dragline silk fibers and spinning conditions, four samples were made at the reeling rates of 1mm/s, 20mm/s, 43.5mm/s and 110mm/s from the major ampullate glands of Araneus Ventricosus and the other two of dragline silks were prepared from a crawling or dropping spider. The surface microstructure and nanofibril characteristic were analyzed with atomic force microscopy (AFM). AFM images of 2000nm*2000nm and 500nm*500nm of these samples showed that the spinning condition influenced the surface roughness and fibril size, while AFM images of 200nm*200nm clearly displayed that dragline silk of Araneus Ventricosus included sheet macro-conformation structure. These results can facilitate the further investigation of the spinning mechanism of a spider in order to understand mechanical properties and macromolecular structures of dragline silk.     

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。     

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.     

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.     

闽北3种人工林土壤游离氨基酸组成及其差异研究

3种人工林土壤游离氨基酸总含量表现出明显的垂直分布特征.针阔叶混交林和针叶林表层(0～20 cm)土壤游离氨基酸含量显著高于深层(20～40 cm)土壤的含量(P＜0.05);而阔叶林表层土壤游离氨基酸含量低于深层土壤的含量,但差异不显著.土壤游离氨基酸各组分均表现出显著的垂直分布特征,阔叶林表层土壤中天冬氨酸、谷氨酸、组氨酸、精氨酸和苯丙氨酸的含量显著高于深层土壤对应组分的含量,但苏氨酸、脯氨酸、甘氨酸、丙氨酸、亮氨酸显著低于深层土壤对应组分的含量;针阔叶混交林表层土 壤中酪氨酸、组氨酸和脯氨酸低于深层土壤的含量,而其余组分均显著高于深层土壤的含量;而针叶林中除甲硫氨酸和赖氨酸外,其余15种氨基酸在表层土壤中的含量均高于深层土壤中对应组分的含量.3种人工林中无论是表层土壤还是深层土壤,游离氨基酸均以中性氨基酸含量为最高,碱性和酸性氨基酸含量次之,含硫氨基酸含量最低.表层土壤中天冬氨酸、丝氨酸、谷氨酸、甘氨酸、丙氨酸、缬草氨酸、甲硫氨酸、异亮氨酸、亮氨酸和精氨酸的含量依次为:针叶林＜阔叶林＜针阔叶混交林,而赖氨酸和脯氨酸的含量则以针叶林为最高,针阔叶混交林次之,阔叶林最低;阔叶林表层土壤中络氨酸和胱氨酸含量最高.深层土壤中各组分氨基酸除赖氨酸之外均以针叶林中含量最低,针阔叶混交林次之,而阔叶林中含量最高.     

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。     

磷酰化氨基酸的特性及应用前景

综述了磷酰化氨基酸的研究进展．磷酰化氨基酸能够发生自身活化成肽、成酯、磷上酯交换和磷酰基的N→0迁移等化学反应，对解释一些生命现象和探讨生命起源具有重要的作用，同时，在生物技术、医药等领域也展示了广阔的应用前景、