摘要
牵引丝(dragline silk)由主壶腹腺蛛丝蛋白(major ampullate spidroin,MaSp)组成,是蜘蛛丝中强度最好的丝,同时具有极佳的生物相容性和可降解性,因此引起研究者的研究热潮。目前关于大腹园蛛MaSp结构和成丝机理方面的研究甚少,限制了其仿生应用。本文以大腹园蛛牵引丝的组成蛋白质之一MaSp1为研究对象,通过锚定PCR的方法首次获取了大腹园蛛MaSp1 NT的完整编码基因,并对其进行了克隆、表达、纯化,产量可达60 mg/L;同时对该MaSp1的CT进行表达纯化,产量可达80 mg/L。另外,通过CD色谱分析了MaSp1 NT和CT的二级结构,结果表明二者的二级结构均以α-螺旋为主。上述结果为大腹园蛛MaSp1的结构和成丝机理研究奠定了基础。
Dragline silk is the strongest of all the spider silks,with superior biocompatibility and biodegradability,which makes it a hot area in biomaterial research.The inadequate understanding of the Araneus ventricosus(A.v.)major ampullate spidroin(MaSp)structure and fiber formation mechanism restricts its bionic application.Herein,A.v.MaSp1,one of the main components of dragline silk,was studied.The full-length N-terminal(NT)coding sequence of A.v.MaSP1 was obtained by anchored PCR,and through gene cloning,expression and purification,the NT protein was harvested at a yield of 60 mg/L.Meanwhile,the C-terminal(CT)of MaSp1 was also expressed and purified with a yield of 80 mg/L.The secondary structures of MaSp1 NT and CT were determined from CD spectra and the curves showed thatα-helix is the main structure of both proteins.The results set the foundation for the research of structure and fiber formation mechanism of A.v.MaSp1.
作者
贾秋品
温睿
李雪
孟清
IA Qiu-pin;WEN Rui;LI Xue;MENG Qing(Institute of Biological Science and Biotechnology,Donghua University,Shanghai 201620,China)
出处
《生命科学研究》
CAS
CSCD
2019年第3期200-207,共8页
Life Science Research
基金
国家自然科学基金资助项目(31570721)
中央高校基本科研业务费专项资金资助项目(CUSF-DH-D-2015045)
