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以羽衣甘蓝(Brassica oleracea var.acephala)自交不亲和系(S13-bS13-b)为试材,从柱头中分离Exo70A1基因,命名为BoExo70A1,GenBank登录号为JF919716。BoExo70A1全长cDNA序列为2 184 bp,包含56 bp的5′非编码区,211 bp的3′非编码区和一个长度为1 917 bp的开放读码框(ORF),对应编码一个含有638个氨基酸残基的蛋白质。氨基酸序列比对分析表明,羽衣甘蓝BoExo70A1与油菜BnExo70A1、拟南芥AtExo70A1的一致性分别为99%和94%;BoExo70A1基因结构含12个外显子和11个内含子,内含子5′供体位点和3′受体位点边界序列符合GU-AG规则;Southern杂交结果显示,BoExo70A1在羽衣甘蓝基因组中存在多拷贝;Northern杂交结果显示BoExo70A1在茎、叶、花瓣、花药、柱头、花柱和子房中表达,而且在叶中的表达量最低。
The Exo70A1 gene was isolated from the stigma of Brassica oleracea var. Acephala self-incompatible (S13-bS13-b) and named as BoExo70A1. The GenBank accession number was JF919716. The full-length cDNA sequence of BoExo70A1 is 2 184 bp in length and contains 56 bp 5 ’non-coding region, 211 bp 3’ non-coding region and 1 917 bp open reading frame (ORF) Amino acid residues of the protein. Amino acid sequence alignment analysis showed that the identity of BoExo70A1 in kale was 99% and 94% respectively in Arabidopsis BnExo70A1 and Arabidopsis AtExo70A1. The structure of BoExo70A1 gene consisted of 12 exons and 11 introns with 5 ’ The results of Southern blot showed that there were multiple copies of BoExo70A1 in the genome of Kale; the results of Northern blot showed that BoExo70A1 was expressed in stem, leaf, petal, anther, stigma, style And ovary expression, and the lowest expression in the leaves.