Cloning，Expression Analysis and Vector Construction of CkCDPK Gene from Caragana korshinskii

doi:10.7668/hbnxb.201751471

Abstract

Abstract: Calcium dependent protein kinase is a key signal transductor, and plays an important role in plant resistance to envirenment stress by regulating and participating in metabolic pathways of plants. To explore the related function genes of caragana, we isolated an open reading frame of CDPK gene from caragana by homology cloning strategy, termed CkCDPK. Bioinformatic analysis showed that the gene had a 1 710 bp open reading frame(ORF), encoding 570 amino acids, with a molecular weight of 64.03 ku and PI of 9.12. Structural analysis showed that the CkCDPK gene contained the N-terminal variable region, a protein kinase domain, EF hand structure, calmodulin domains, etc. Quantitative real-time PCR technique was used to detect CkCDPK expression level under adverse situations. The results revealed that the CkCDPK presented a unimodal trend after treated with salinity(NaCl) and drought(PEG), of which the gene expression levels were higher at 6 h of salinity stress and 4 h of drought stress. Under ABA stress, the gene expression was up-regulated compared with the contrast. These results indicated that the CkCDPK gene might be involved in the regulation of environmental stress responses in Caragana korshinskii. The plant expression vector pCAMBIA3301-CkCDPK was also constructed. This study provided the theoretic and experimental basis for further study of the CkCDPK function in stress-tolerance regulation in Caragana korshinskii.

Key words: Caragana korshinskii, Calcium-dependent protein kinases(CDPK), Adversity stress, Vector construction

CLC Number:

Q78
S793.3

JIA Huili, GUO Jicheng, WU Xinming, WANG Yunqi, LIU Jianning, FANG Zhihong, SHI Yonghong, DONG Kuanhu. Cloning，Expression Analysis and Vector Construction of CkCDPK Gene from Caragana korshinskii[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 43-51. doi: 10.7668/hbnxb.201751471.

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