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以茎瘤芥(Brassica juncea var.tumida Tsen et Lee)‘永安’为材料,通过RACE和RT-PCR技术得到1个AP2/EREBP转录因子基因的cDNA全长序列和基因组DNA(gDNA)序列,该基因与拟南芥AtABR1基因的氨基酸序列相似性高达72%,因此命名为BjABR1(GenBank登录号:JQ713825.1)。BjABR1基因gDNA序列含1个内含子;cDNA序列全长1 514 bp,含有1个1 146 bp的开放阅读框(ORF),编码381个氨基酸;其推定编码的蛋白分子量为41.674 kD,等电点为9.11,具有14个磷酸化位点,含1个典型的AP2 DNA结合域和1个CMX-1基序。洋葱表皮细胞的瞬时表达显示,BjABR1蛋白定位于细胞核。荧光定量PCR分析结果表明,该基因在茎瘤芥不同发育时期的根、茎、叶中均有表达,但在根中表达量极高;在对茎瘤芥组培幼苗进行高盐、渗透压和低温3种非生物胁迫处理后发现,3种胁迫均能诱导该基因表达,但其对高盐的响应更为迅速。
The full-length cDNA and genomic DNA sequences of one AP2 / EREBP transcription factor gene were obtained by RACE and RT-PCR using ’Yong’an’ (Brassica juncea var. Tumida Tsen et Lee) The deduced amino acid sequence of Arabidopsis AtABR1 was 72% similar to that of Arabidopsis thaliana, and was named BjABR1 (GenBank accession number: JQ713825.1). The gDNA sequence of BjABR1 gene contains 1 intron. The full-length cDNA of BjABR1 gene is 1 514 bp in length and contains a 1 146 bp open reading frame (ORF) encoding 381 amino acids. The deduced protein has a molecular weight of 41.674 kD and an isoelectric point Point was 9.11 with 14 phosphorylation sites containing 1 typical AP2 DNA binding domain and 1 CMX-1 motif. Transient expression of onion epidermal cells showed that BjABR1 protein localized in the nucleus. Fluorescent quantitative PCR analysis showed that the gene was expressed in roots, stems and leaves at different developmental stages of the tumorous stem mustard, but its expression was extremely high in the root. In the tissue culture of the tumorous stem mustard, high salt and osmotic pressure After three kinds of abiotic stress treatments, the three kinds of stress could induce the gene expression, but its response to high salt was more rapid.