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      Characterization and Expression Analysis of Protein Kinase C Gene from Dunaliella salina

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          Abstract

          Protein kinase C (PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene from Dunaliella salina ( DsPKC) using rapid amplification of cDNA ends (RACE) and RT-PCR methods. And we submitted the mRNA sequence of DsPKC gene to NCBI (Genbank No. JN625213). In the present paper, the DsPKC gene open reading frame obtained by PCR was cloned into pGS-21a vector and transformed into Escherichia coli to generate the fusion protein. Bioinformatics analysis revealed that DsPKC gene was a member of serine/threonine kinase with two conserved domains and highly conserved motifs. The DsPKC was highly expressed upon induction with isopropyl-β- d-thiogalactoside (IPTG) at a final concentration of 0.2 mmol L −1 at 37℃. Under salt stress, the fusion protein Green Fluorescent Protein (GFP)-DsPKC was transferred from the cytoplasm to the cell membrane. The expression pattern of DsPKC gene was analyzed using real-time quantitative PCR, and indicated that DsPKC gene was up-regulated by 3.0 mol L −1 NaCl at 12 h, which was significantly higher than in control values ( P < 0.05). These results suggest that the DsPKC gene plays an important role in response to salt stress in D. salina.

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          Author and article information

          Journal
          JOUC
          Journal of Ocean University of China
          Science Press and Springer (China )
          1672-5182
          06 July 2019
          01 October 2019
          : 18
          : 4
          : 977-984
          Affiliations
          1Key Laboratory of Hydrobiology in Liaoning Province’s University, Dalian Ocean University, Dalian 116023, China
          2Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea, Ministry of Agriculture, Dalian Ocean University, Dalian 116023, China
          Author notes
          *Corresponding author: CHAI Xiaojie
          Article
          s11802-019-3799-4
          10.1007/s11802-019-3799-4
          Copyright © Ocean University of China, Science Press and Springer-Verlag GmbH Germany 2019.

          The copyright to this article, including any graphic elements therein (e.g. illustrations, charts, moving images), is hereby assigned for good and valuable consideration to the editorial office of Journal of Ocean University of China, Science Press and Springer effective if and when the article is accepted for publication and to the extent assignable if assignability is restricted for by applicable law or regulations (e.g. for U.S. government or crown employees).

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          Self URI (journal-page): https://www.springer.com/journal/11802

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