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目的:对前期已经克隆的丹参SmERF1基因进行聚类分析,分析SmERF1基因在不同诱导子处理后不同时间的表达情况;并对其进行亚细胞定位分析。方法:利用MEGA5软件对SmERF1基因及拟南芥ERF基因家族进行聚类分析;利用半定量RT-PCR方法,分析SmERF1基因在不同诱导子处理后不同时间的表达情况;将SmERF1与GFP融合,在洋葱表皮瞬时表达,以确定SmERF1蛋白表达部位。结果:丹参SmERF1属于ERF家族第Ⅶ亚族;YE+Ag+诱导对SmERF1基因的表达没有影响,ABA和MeJA可以抑制该基因的表达,而水杨酸诱导会出现先抑制,后随处理时间加长,SmERF1基因表达又恢复;亚细胞定位确定SmERF1基因在细胞核内特异表达。结论:SmERF1是一个AP2/ERF转录因子,属于AP2/ERF家族第Ⅶ亚族,其表达受ABA,SA,MeJA等激素调节控制。
OBJECTIVE: To cluster the SmERF1 gene of Salvia miltiorrhiza which had been cloned in the early stage and analyze the expression of SmERF1 gene at different time points after different inducer treatments. The subcellular localization of SmERF1 gene was also analyzed. Methods: SmERF1 gene and Arabidopsis ERF gene family were clustered by using MEGA5 software. The expression of SmERF1 gene at different time after different elicitors was analyzed by semi-quantitative RT-PCR. SmERF1 and GFP were fused in Onion epidermis transient expression to determine SmERF1 protein expression sites. Results: SmERF1 of Salvia miltiorrhiza belongs to the Ⅶ subfamily of ERF family. The induction of YE + Ag + had no effect on the expression of SmERF1 gene. ABA and MeJA could inhibit the expression of SmERF1, while the salicylic acid induced inhibition first, followed by longer treatment time, SmERF1 gene expression and recovery; subcellular localization of SmERF1 gene in the nucleus specific expression. Conclusion: SmERF1 is an AP2 / ERF transcription factor belonging to the Ⅶ subfamily of AP2 / ERF family. Its expression is regulated by ABA, SA, MeJA and other hormones.