Clone and expression of calcium-dependent protein kinase BdCDPK14 gene from Brachypodium distachyon

ObjectiveThe present experiment was conducted to clone calcium dependent protein kinase (CDPK) Bd-CDPK14 gene from Brachypodium distachyon and detect its expression level under drought stress, in order to provide a foundation for further research on regulation mechanism of BdCDPK14 gene in response to drought. MethodSpecific primers were designed based on NCBI retrieval results, and cDNA of B. distachyon was used as template. BdCDPK14, one of B. distachyon CDPK genes was cloned by PCR amplification. Encoded protein of BdCDPK14 gene was analyzed by online software on its bioinformatics. Its expression level under PEG-6000 drought stress was detected by RT-PCR. ResultBd-CDPK14 gene(GenBank accession number: XM_003564390) cloned from B. distachyon was 1750 bp in length, open reading frame(ORF) was 1545 bp CDS, coding 514 amino acids. The molecular weight of encoded protein was 56.78 kD, isoelectric point 5.45, fat index 78.21 and instability index 38.66, belonging to stable protein. BdCDPK14 protein con-tained several domains including four EF-hands, protein tyrosine kinase domain, lipopolysaccharide kinase family, ATP-binding region, serine/threonine protein kinase active-site region and predicted transmembrane region. It shared the closest genetic relationship with TaCPK13 protain(ABY59018) of wheat. BdCDPK14 protein was a protein with random coil andα-helix located in chloroplast and plasma membrane. ConclusionThe cloned BdCDPK14 gene from B. distachyon leaf is a member of CDPK genes, and is involved in drought-resistant reaction in B. distachyon. So it can be served as a candi-date gene to further study its role in drought-resistant in B. distachyon.