论文部分内容阅读
以菜薹[Brassica rapa L.ssp.chinensis(L.)Mokino var.utilis Tsen et Lee]为材料,克隆得到1个WRKY转录因子,命名为BrWRKY75。氨基酸序列比对及进化树分析发现BrWRKY75与拟南芥的WRKY75同源性较高,且同属于第Ⅱ类c亚族。实时荧光定量PCR表明BrWRKY75在菜薹叶片衰老过程中表达明显增强。亚细胞定位和转录活性分析显示BrWRKY75定位于细胞核,是一种核蛋白,并且具有转录抑制活性。体外凝胶阻滞试验(EMSA)证实Br WRKY75能与W-box(TTGAC)元件特异结合。上述结果为进一步研究菜薹叶片衰老的转录调控机制奠定了基础。
A WRKY transcription factor named BrWRKY75 was cloned from Brassica rapa L.ssp.chinensis (L.) Mokino var. Utilis Tsen et Lee. Amino acid sequence alignment and phylogenetic tree analysis showed that BrWRKY75 had high homology with WRKY75 in Arabidopsis thaliana, belonging to the second subgroup c subfamily. Real-time quantitative PCR showed BrWRKY75 expression significantly increased during the senescence of cauliflower leaves. Subcellular localization and transcriptional activity analysis revealed that BrWRKY75 is localized to the nucleus, a nuclear protein, and has transcriptional inhibitory activity. In vitro gel blocking assay (EMSA) confirmed that Br WRKY75 specifically binds to the W-box (TTGAC) element. These results laid the foundation for the further study of the transcriptional regulation mechanism of the senescence of cauliflower leaves.