作物学报2012，Vol.38Issue(4)：578-588,11.DOI:10.3724/SP.J.1006.2012.00578
甘蓝、大白菜和甘蓝型油菜EXO70A1基因的克隆与表达特性
Cloning and Expression Characteristics of EXO70A1 from Brassica oleracea,Brassica campestris, and Brassica napus
摘要
Abstract
EXO70A1 is an important signal element of self-incompatibility in Brassica. In this study, coding sequences (CDS) of EXO70A1 and the corresponding gDNA were cloned by PCR. Sequence analysis was carried out by means of bioinformatics. RT-PCR was used to analyze expression characteristics of EXO70A1 in Brassica oleracea. And BoEXO70Al was transformed into yeast Y187 to explore its expression. The results showed: That the gene lengths of BnEXO70Al, BrEXO70Al and BoEXO70A1 were 3 797, 3 752, and 3 770 bp, respectively, and all consisted of 12 exons and 11 introns; their identity positions reached 91%. Sequence conservation was higher in exons than in introns excluding 4th, 5th, 6th, and 8th intron. CDS of three EXO70A1 genes consisted of 1 917 base pairs and sequence similarity was 97.1%. Three proteins of EXO70A1 conduced all consisted of 638 amino acids; the similarity and identity of EXO70A1 in the three species were 99.8% and 98.1%, respectively. BnEXO70A1, BrEXO70A1, and BoEXO70A1 shared similar secondary structures and three-dimensional structures. All introns started from the sequence GU and ended with the sequence AG (in the 5' to 3' direction). They are referred to as the splice donor and splice acceptor site, respectively. Another important sequence "CU(A/G)A(C/U)" in all introns of EXO70A1 genes was located at 20-50 bases upstream of the acceptor site. The corresponding sequence lengths of all 12 exons of EXO70A1 were identical among three Brassica species and Arabidopsis thaliana; and similarity of four coding sequences was 90.1%. The similarity and identity of EXO70A1 proteins in the four species reached 99.8% and 93.7%, respectively. EXO70A1 was a subunit of EXO70 family which showed high conservative in Magnoliophyta. The results showed that BoEXO70Al presented weak expression in Y187, while expressed in stamens, young stems, petals, pistils, young roots and leaves. However, its expression quantity was different in different tissues, with strong expression in pistils and weak in stamens. Thus conclusions can be drawn that: EXO70A1, in Brassica, is high conservative; EXO70A1 perhaps plays very important roles in plant cells because of its different expression characteristics in different plant tissues.关键词
EXO70A1/克隆/序列分析/表达特性Key words
EXO70A1/Cloning/Sequence analysis/Expression characteristics引用本文复制引用
杨昆,高启国,周永祥,张贺翠,赵永斌,杨永军,陆俊杏,朱利泉,薛丽琰,吕俊..甘蓝、大白菜和甘蓝型油菜EXO70A1基因的克隆与表达特性[J].作物学报,2012,38(4):578-588,11.基金项目
本研究由中央高校基本科研业务费专项资金(XDJK2011C012,XDJK2012B18,XDJK2009C 110),高等学校学科创新引智计划(B12006),国家自然科学基金项目(30971849,3090986)和重庆市自然科学基金项目(2012jjB014,2009bb1298)资助. （XDJK2011C012,XDJK2012B18,XDJK2009C 110）
