悦目金蛛丝蛋白TuSp1重复模块特征  

Characterization of Spidroin TuSp1 Repeat Module from Argiope Amoena

作　　者：王金雷[1] 陈格飞[1] 李佳[1] 林瑛[1] 孟清[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620

出　　处：《东华大学学报（自然科学版）》2013年第2期196-201,200-201,共6页Journal of Donghua University(Natural Science)

基　　金：国家"八六三"高技术研究发展计划资助项目(2006AA03Z451);国家自然科学基金资助项目(31070698);上海市基础研究重点资助项目(10JC1400300);教育部高等学校博士学科点专项科研基金资助项目(20120075110007)

摘　　要：针对蜘蛛管状腺丝蛋白(TuSp)结构进行克隆和分析,基于悦目金蛛(Argiope amoena,A.amoena)管状腺总RNA,利用简并引物和巢式聚合酶链反应(PCR)技术克隆AaTuSp1编码基因序列1273bp,其包含3个重复单元,每个重复单元编码176个氨基酸,重复单元之间的序列同源性高达97%以上,且富含丙氨酸(Ala)和丝氨酸(Ser),其次是甘氨酸(Gly),组成了一系列PolyT,PolyA,PolyS,SQ和GX模体结构,推测与其独特的力学性能相关.系统进化分析表明,不同于典型蛛丝蛋白MaSp,Flag等,AaTuSp1及近系包卵丝蛋白形成一个独立的进化分支.研究结果为基于TuSp1合成仿生蛛丝提供了新的结构基因蓝图.Cloning and structural analysis of tubuliform spidroin （TuSp） were performed based on the extraction of total RNA of Argiope amoena tubuliform, degenerate primers and the technique of nested polymerase chain reaction（PCR）were used to obtain AaTuSpl, which was 1 273 basepairs, and included three repeats. Each repeat contained 176 amino acids, and among these repeats, the identity was up to 97%. The sequence was rich in Ala, Ser and Gly, and formed a series of motifs, such as PolyT, PolyA, PolyS, SQ and GX, which may be related with the mechanical characterization. Phylogenetic analysis demonstrated that AaTuSpl formed a new branch that was different from MaSp, Flag and so on. The research results provided a new gene blueprint for biomimicing spider silk.

关 键 词：悦目金蛛 管状腺丝蛋白 重复单元 巢式聚合酶链反应(PCR) 进化 

分 类 号：Q78[生物学—分子生物学]