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Expression, purifi cation, and subcellular localization of phospholipase C in Dunaliella salina

  • Yuting Cong
  • Yuan Wang
  • Jinrong Yue
  • Zhenyu Xing
  • Xiangnan Gao
  • Xiaojie ChaiEmail author
Article
  • 3 Downloads

Abstract

Plants possess eff ective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specifi c phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purifi cation, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purifi ed and identifi ed using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fl uorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced signifi cantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress.

Key word

Dunaliella salina DsPLC gene prokaryotic expression subcellular localization salt stress 

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Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Yuting Cong
    • 1
  • Yuan Wang
    • 1
  • Jinrong Yue
    • 1
  • Zhenyu Xing
    • 1
  • Xiangnan Gao
    • 1
  • Xiaojie Chai
    • 1
    Email author
  1. 1.Key Laboratory of Hydrobiology in Liaoning Province, College of Fisheries and Life ScienceDalian Ocean UniversityDalianChina

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