生命科学研究2017，Vol.21Issue(5)：392-398,416,8.DOI:10.16605/j.cnki.1007-7847.2017.05.004
大腹园蛛鞭状腺蛛丝蛋白N端结构域的克隆表达及圆二色谱分析
Cloning and Expression and CD Spectra of the N-terminal Domain of Flagelliform Spidroin from Araneus ventricosus
摘要
Abstract
The flagelliform spider silk,which is composed of the flagelliform spidroin (FlSp),is the most extensible among the six spider silks.It has wide potential applications.However,few sequences of the terminal domains of FlSp have been reported and their structures and functions are also ambiguous.These problems have limited the application of the flagelliform spider silk.Here,the N-terminal domain of FlSp from Araneus ventricosus (FlSp_A.v-NT) was obtained via the 5'-RACE method based on the genomic RNA and DNA.The FlSp_A.v-NT was further successfully cloned,expressed and purified with the final yield of 60 mg/L.Meanwhile,for the first time,the secondary structure of FlSp NT domain was analyzed using circular dichro ism (CD) spectra.It indicated that the α-helix was the predominant secondary structure of FlSp_A.v-NT.These results provide research material and useful information for NT structural and functional analysis of FlSp.关键词
大腹园蛛/蛛丝蛋白/5'-RACE方法/FlSp_A.v-NT编码基因/克隆表达/二级结构Key words
Araneus ventricosus/spidroin/5'-RACE/FlSp_A.v-NT coding gene/cloning and expression/secondary structure分类
生物科学引用本文复制引用
李雪,陈格飞,张晓玲,孟清..大腹园蛛鞭状腺蛛丝蛋白N端结构域的克隆表达及圆二色谱分析[J].生命科学研究,2017,21(5):392-398,416,8.基金项目
国家自然科学基金资助项目(31570721) （31570721）
教育部直属高校学校特色项目(TS2011DHDX025) （TS2011DHDX025）
上海市科委攻关项目(14521100700) （14521100700）
上海市科委国际合作项目(14520720200) （14520720200）
中央高校基本科研业务费专项资金资助项目(17D311308) （17D311308）
