关键词: 羽衣甘蓝, 自交不亲和性, ARC1, Exo70A1, 结构域

Abstract: The yeast-two hybrid system was used to detect the interaction between ARC1 and Exo70A1 from ornamental kale（Brassica oleracea var. acephala）. The nucleotide sequences of full-length BoARC1，N terminal of BoARC1（1–279 aa，UND domain），C terminal of BoARC1（280–663 aa，U-box domain and Arm repeat domain）were constructed into the vector pGBKT7，respectively. Thus，the plasmids BD-ARC1，BD-ARC1-N or BD-ARC1-C were obtained and confirmed. On the other hand，the nucleotide sequences of full-length BoExo70A1，BoExo70A1-Δ1（1–85 aa），Exo70A1-Δ2（1–270 aa），Exo70A1-Δ3（271–638 aa，Exo70 domain）were constructed into the vector pGADT7，respectively. And the plasmids AD-Exo70A1，AD-Exo70A1-Δ1，AD-Exo70A1-Δ2 or AD-Exo70A1-Δ3 were also obtained and confirmed. These plasmids were used to co-transform yeast strain Y187，which were then plated onto SD/-Leu-Trp（synthetic dropout glucose agar plates lacking leucine and tryptophan）. The β-galactosidaseassays showed that yeast cells co-transformed with the BoARC1-N/AD-Exo70A1-Δ1 plasmids or BoARC1-N/BD-Exo70A1-Δ2 plasmids turned blue. These results indicated that N terminal of BoARC1 and BoExo70A1 mediated interaction between BoARC1 and BoExo70A1.

Key words: Brassica oleracea var. acephala, self-incompatibility, ARC1, Exo70A1；domain