Abstract:Abstract: The rice FERONIA-like receptor 1 (FLR1) is a member of receptor-like kinase involving in rice development and stress resistance. Bioinformatics predicted that FLR1 protein can be divided into N-terminal (FLR1-BW, 1~450 aa) and C-terminal (FLR1-BN, 474~892 aa) domains. To elucidate the function of FLR1, its N-terminal and C-terminal domains were cloned into expression vectors pET-32a, pGEX-4T-1, pET-22b, and pTriEx-4, respectively. The plasmids were transformed into BL21 (DE3) and proteins were expressed under different conditions. The results showed that FLR1-BW was successfully expressed in E. coli from pET-32a and pGEX-4T-1 and the optimal conditions were 16 ℃/16 h with 0.5 mmol/L IPTG. The recombinant FLR1-BW was purified to homogeneity and used for antibody production in rats. However, FLR1-BN protein was cytotoxic to E. coli, but was expressed successfully in 293 cells. The research put forward a new strategy for receptor-like kinase antibody preparation and laid a foundation for studying the functions of FLR1 in the future.
引用本文:
吴秀秀, 蒋 顺, 于 峰, 曹建中, 汪 龙. 水稻中类受体蛋白激酶FERONIA-like Receptor 1的表达纯化及抗体制备[J]. 生命科学研究, 2019, 23(5): 393-400.
WU Xiu-xiu, JIANG Shun, YU Feng, CAO Jian-zhong, WANG Long. Expression, Purification and Antibody Preparation of Rice FERONIA-like Receptor 1. Life Science Research, 2019, 23(5): 393-400.