大腹圆蛛Fosmid文库的构建及MaSp基因筛选方法的探讨

介绍了构建大腹圆蛛Fosmid基因组文库及牵引丝蛋白(MaSp)基因克隆筛选的全过程.采用改良CTAB法提取大片段基因组DNA,通过自主构建的电洗脱核酸回收装置分离回收30～40kbDNA片段,经补平磷酸化、与pCC2FOS载体连接、体外包装和转染EPI300TM-T1R,首次构建了无偏向性的大腹圆蛛Fosmid文库,其滴度为4.5×105cfu/mL,覆盖基因组倍数为10.以α-32P标记寡核苷酸探针对文库进行初步筛选,获得含MaSp基因的12个阳性克隆.该文库符合Fosmid文库的品质要求,为进一步筛选并研究大腹圆蛛MaSp基因序列奠定了基础.     

大腹园蛛Avg1 cDNA的克隆和序列分析

组织蛋白酶(cathepsin)是一大类裂解肽键的蛋白水解酶,根据其催化中心不同,组织蛋白酶B(CB)属于半胱氨酸蛋白酶,易被巯基试剂抑制,因此又是巯基酶类,隶属于木瓜蛋白酶族[1 ] .CB可分解胶原蛋白、层粘连蛋白等基底膜的主要成分,近年来发现与癌细胞的浸润和转移有密切的关系[2 ]     

A derivative of the orb web and its evolutionary significance

Abstract

This paper describes a new kind of ladder-web structure in which there are two ladders, one above and one below a centrally positioned orb. It differs from previously described ladder-webs, not only because of the two ladders but also because of its 24 h (or more) duration, its vertical placement against the trunks of trees, and the fact that it apparently offers the spider protection against parasitism. Both the spider (Araneus atrihastulus) and its ladder-web are ideally adapted to the tree-trunk: the web with regard to its position, shape, and lack of visibility; and the spider in respect of its coloration, daytime posture, and proximity to the snare. It is concluded that the design of this web offers a number of advantages which evidently enhance the spider's survival and increase its capture potential over and above that of the simple orb.     

中国园蛛科1新种和2个新雄性(蜘蛛目:园蛛科)

本文描述了园蛛科3种蜘蛛：多齿园蛛,新种Araneus polydentatus sp．nov．,舜皇园蛛Araneus shunhuangensis Yin et al,1990和松阳芒果蛛Mangora songyangensis Yin et al,1990。舜皇园蛛和松阳芒果蛛的雄性均系首次报道。本文同时提供了这3种蜘蛛的地理分布图。     

中国六盘山园蛛属和吉园蛛属二新种记述(蜘蛛目,园蛛科)(英文)

描述采自宁夏六盘山国家级自然保护区的园蛛科2新种:镰状园蛛Araneus falcatus sp. nov.和叉吉园蛛Gibbaranea bifida sp.nov.。模式标本保存于河北大学博物馆(MHBU)。     

大腹园蛛鞭毛样丝蛋白cDNA克隆

应用RT-PCR技术,从大腹圆蛛（Araneus ventricosus)壶腹腺中扩增出鞭毛样丝蛋白基因（flagelldid-form silk protein gene),经1.5%琼脂糖凝胶电泳分离,WizardPCR Preps DNA Purification System回收后,将其克隆在pGEM-T载体中,经限制性核酸内切酶鉴定和核苷酸序列分析证实,构建的重组擀粒pSF1中含有蜂蛛鞭毛样丝蛋白基因,且含有3个重复序列。     

大腹园蛛丝腺及其细胞形态的初步观察

运用显微解剖和石蜡切片技术,我们首次对大腹园蛛Araneus ventricosus腹部的5种丝腺的形态进行了分离、拍照和切片观察.研究发现壶状腺腹腔较大,有多根纺管;梨状腺为多细胞腺体、成簇;集合腺有不规则的分支,纺管外附着小结节;葡萄状腺成簇.各类型丝腺腺体细胞明显可见,但未能明显见到不同丝腺腺体细胞的形态区别.同时也发现了几个存疑形态：壶状腺上附着的腺体、壶状腺下导管和壶状腺腺体细胞,它们可能与蜘蛛腺体的退化或发育等相关.     

大腹园蛛拖丝蛋白一级结构初步研究

采用部分酸水解的方法对大腹园蛛（Ａｒａｎｅｕｓｖｅｎｔｒｏｃｏｓｕｓ）拖丝纤维蛋白进行水解,通过反相高液相色谱分离到一些重复小肽片段,对其序列分析表明,该蜘蛛拖丝蛋白与肖蛸科的棒络新妇蛛（Ｎｅｐｈｉｌａｃｌａｖｉｐｅｓ）丝蛋白的序列比较,有一个完全相同的肽段ＧＹＧＰＧ,其余所测片段测存在差异,显示不同种属的蜘蛛丝在一级结构上的同异,并探讨了结构和功能的关系。     

Silk Production after Mechanical Pulling Stimulation in the Ampullate Silk Glands of the Barn Spider, Araneus cavaticus

Synthesis of protein by the major ampullate silk glands in the barn spider, Araneus cavaticus was stimulated by depleting the storage of silk protein in the ampulla by mechanically pulling fiber from the spigot. After this treatment, fine structural changes of the glandular epithelium during silk production were examined using light and transmission electron microscopes. In the process of rapid production, major secretory silk was synthesized at the tail region via rER of glandular epithelial cells, and was transported into the ampulla region. The mature secretory product in glandular epithelium appears almost spherical vacuoles which were grown up by fusion with the surrounding small vesicles including the secretory silk. Unlike to a typical process of the secretion, the ampullate silk of tail region seems to bypass either concentrating or packaging steps by the Golgi apparatus. However there's no doubt that the Golgi apparatus also play an important role in the secretory process of the ampulla region. After mechanical pulling stimulation, both epithelia of ampulla and tail regions appeared as a thinner layer of columnar cells with less definitive cell membrane. There are few secretory droplets within these cells, thus causing this region to stain much lighter. It is obvious that the cell loses part of its cytoplasm in this process, and disorganization of the secretory product occurs when it is extruded from the cells by a apocrine release.