[Cloning and bioinformatic analyzing of transcription factor AP2/ERF-B3 subfamily genes from Brassica napus L. Huyou 15].

AP2/ERF is a large family of transcription factors in plant.Genes in the AP2/ERF family encode transcriptional regulators with a variety of functions involved in the developmental and physiological processes in plants.Two AP2/ERF family transcriptional regulators(BnaERFB3-1 and BnaERFB3-2)were isolated from B.napus by in silico cloning method using the conserved domain amino acid sequence of A.thaliana AP2/ERF-B3 subfamily as probe.Based on the sequences of BnaERFB3-1 and BnaERFB3-2,we isolated the BnaERFB3-1-Hy15 gene and BnaERFB3-2-Hy15 gene from winter and spring type B.napus L.cv Huyou15 by RT-PCR and PCR using cDNA and DNA as template.DNA sequencing and analyzing indicated that there was only one amino acid residue difference between BnaERFB3-1 and BnaERFB3-1-Hy15,BnaERFB3-2 and BnaERFB3-2-Hy15,respectively.No intron localized on the two genes from Huyou15.Then,deduced amino acid sequence,composition,hydrophobicty and hydrophilicity,physical and chemical characterization,phylogenetic tree,conserved domain sequences,function domain,molecular modeling,and folding state were predicted and analyzed.BnaERFB3-1-Hy15 and BnaERFB3-2-Hy15 were hydrophilic protein.The two proteins and AtERF5 have similar three-dimension structure.The disordered residues of protein BnaERFB3-1-Hy15 and BnaERFB3-2-Hy15 were higher than that of AtERF5.BnaERFB3-1 was mainly expressed in seed,while BnaERFB3-2 was mainly expressed in root.Moreover,those genes were successfully constructed into the recombinant plasmids of plant expression vector and yeast expression vector,which established a base for transformation of oilseed and studies of those genes function in abiotic stresses of B.napus.