天津科技大学学报2024，Vol.39Issue(6)：14-20,7.DOI:10.13364/j.issn.1672-6510.20230193
莱茵衣藻小G蛋白ARL1的原核表达、纯化及多克隆抗体制备
Prokaryotic Expression,Purification and Polyclonal Antibody Preparation of Small G Protein ARL1 of Chlamydomonas reinhardtii
摘要
Abstract
In order to study the role of small G protein ARL1 in ciliary formation and ciliary signal transduction,a highly specific polyclonal antibody against ARL1 of Chlamydomonas reinhardtii was prepared.First,6×His labeled prokaryotic expression vector pET-28a(+)-arl1 was constructed,and the fusion protein 6×His-ARL1 was induced in E.coli DH5α/C+,and then purified and immunized New Zealand male white rabbits.After three times of immunization,the serum of auricular artery was taken,and the titer was tested by indirect ELISA.ARL1 antibody with high sensitivity and specificity was ob-tained by affinity purification of Protein A and antigenic antibody.By immunofluorescence and Western blot detection,ARL1 was found to be localized in cytoplasm,basal body and cilia,which has laid a foundation for further study on the func-tion of ARL1 in cilia generation and signal transduction of C.reinhardtii.关键词
ARL1/原核表达/多克隆抗体/莱茵衣藻Key words
ARL1/prokaryotic expression/polyclonal antibody/Chlamydomonas reinhardtii分类
生物科学引用本文复制引用
陶冶,鲍冬雪,刘雁霞,樊振川..莱茵衣藻小G蛋白ARL1的原核表达、纯化及多克隆抗体制备[J].天津科技大学学报,2024,39(6):14-20,7.基金项目
国家自然科学基金面上项目(32070698) （32070698）
国家自然科学基金青年科学基金项目(32200558) （32200558）
