关键词: 羽衣甘蓝, 自交不亲和, ARC1, 抗体制备, 蛋白表达

Abstract:

Self-incompatibility is an effective way to avoid inbreeding depression and promote out-crossing in flowering plants. Armadillo Repeat Containing protein 1（ARC1），the downstream factor in SI pathway，can specifically mediated SI pathway. In Brassica oleracea var. acephala，we constructed the full protein（1–663 aa，BoARC1-F），NUD domain（1–279 aa，BoARC1-N）and ARM domain（361–663 aa，BoARC1-C）prokaryotic expression plasmids. SDS-PAGE results showed that the BoARC1-F（68 kD），BoARC1-N（33 kD）and BoARC1-C（38 kD）could be induced by IPTG and purified by affinity chromatography using Ni²⁺-NTA resin. We prepared the monoclonal antibodies against BoARC1-C through immune mouse and cell fusion technology. Cross reaction analysis showed that the monoclonal antibodies could specific react with BoARC1. The expression level of BoARC1 in various tissue and different developmental stigmas were detected. Western blot results revealed BoARC1 mainly expressed in stigma and its expression level reached the maximum at opening flower.

Key words: Brassica oleracea var. acephala, self-incompatibility, ARC1, monoclonal antibody, protein expression