Cloning and Prokaryotic Expression of PsnAP1-1 and PsnAP1-2 Genes in Poplar(Populus simonii × Populus nigra)
Flowering is the transition from vegetative to reproductive phase in plants,and it is regulated by APETALA1 (AP1) gene.We cloned two full-length cDNA sequences of homologous gene AP1 from Populus simonii × Populus nigra by RT-PCR,named as PsnAP1-1 (GenBank No.KC866354) and PsnAP1-2 (GenBank No.KC866355).PsnAP1-1 contains an 726 bp open reading frame (ORF) corresponding to a deduced protein of 241 amino acids,while the estimated molecular weight and the isoelectric point of the putative protein were 28.1 kD and 8.19.PsnAP1-2 with a open reading frame(ORF) of 750 bp,encoding 249 amino acid with a predicted molecular mass of 28.7 kD and a pI of 9.07.A comparison of the deduced amino acid residues indicated that PsnAP1-1 and PsnAP1-2 nucleotide sequences were 71％ and 67％ identities with AP1 gene homologues of Arabidopsis thaliana.With expression analysis by RT-PCR,the PsnAP1-1 and PsnAP1-2 genes only expressed in flower buds,but not expressed in root,leaf,and stem tissues.Furthermore,we constructed recombinant plasmid pET-PsnAP1-1 and pET-PsnAP1-2,transformed to E.coli (BL21),and then induced proteins expression by IPTG.Two 35 kD recombinant proteins were expressed and separated by SDS-PAGE electrophoresis.Our results will provide theoretical and technical bases for analyzing the interaction mechanism of AP1,other MADS-box protein,and the molecular regulation of floral meristem in poplar.
Populus simonii × Populus nigraAPETALA1sequence analysisprokaryotic expression
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