甘蔗转录激活因子ScCBF1基因的克隆与表达分析

Molecular Cloning and Expression Analysis of Transcriptional Activators ScCBF1 Gene from Sugarcane

C-repeat/dehydration-responsive element binding factor (CBF) plays an important role in improving plant stress resistance. The CBF can induce a series of abiotic stress responsive gene expression when the plants are subjected to low temperature, high salt, drought or other abiotic stresses. In this paper, a new CBF-like gene, designed as ScCBF1, was cloned by bioinformatics and RT-PCR method from sugarcane. Sequence analysis showed that ScCBF1 contained a 603 bp open reading frame (ORF) and encoded a deduced protein of 200 amino acids. Its molecular weight and isoelectric point were predicted as 22.80 kD and 10.31, respectively. The amino acid sequence alignment results showed that ScCBF1 shared the similarity of 96% and 94% with the CBF1 protein from Sorghum bicolor and Zea mays, respectively. Phylogenetic tree analysis revealed that ScCBF1 had closer relationships with Sorghum bicolor. qRT-PCR showed that ScCBF1 expressed in root, stem and leaf in sugarcane with the highest expression level in roots. ScCBF1 gene was induced by low temperature, drought or abscisic acid (ABA), but was downregulated under high salinity. The protokaryotic expression vector pGEX-6P-1-ScCBF1 was successfully constructed and transformed into E. coli. Under the induction of IPTG, ScCBF1was successfully expressed in E. coli. This study sheds light on the understanding of the function of ScCBF1.