PJB-2019-981
Cloning and characterization of three cytokinin oxidase/dehydrogenase genes in Bambusa oldhamii
Chun-Yen Hsieh
Abstract
Cytokinin oxidase/dehydrogenase catalyzes the irreversible breakdown of the active cytokinins which are a class of plant hormones that regulate cell division. According to the conserved sequences of CKX genes from monocotyledons, PCR primers were designed to synthesize a probe for screening bamboo genomic library. Cloned result of three genes encoding cytokinin oxidase, were named as followed: BoCKX1, BoCKX2 and BoCKX3. BoCKX1 contains an open reading frame (ORF) of 1,578 bp encoding a polypeptide of 525 amino acid residues with a predicted mass of 57.0 kDa. BoCKX2 contains an ORF of 1,572 bp encoding a polypeptide of 523 amino acid residues with a predicted mass of 57.4 kDa. BoCKX3 contains an ORF of 1,569 bp encoding a polypeptide of 522 amino acid residues with a predicted mass of 56.6 kDa. Comparison the exon-intron structures among the above three genes, there are three exons and two introns in BoCKX1 and BoCKX3 genes, whereas BoCKX2 contains four exons and three introns. The amino acid sequence of BoCKX2 shares 78% and 79% identity with BoCKX1 and BoCKX3, respectively. BoCKX1 and BoCKX3 are particularity closely related that the amino acid and nucleotide sequence identities are more than 90%. These three BoCKX proteins carried putative signal peptide sequences typical of secretion pathway, and a GHS-motif was found at N-terminal FAD binding domain, suggesting that BoCKXs might covalently bind to FAD through a histidine residue.
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