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Molecular cloning and expression analysis of major intrinsic protein gene in Chlamydomonas sp. ICE-L from Antarctica

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Abstract

Major intrinsic proteins (MIPs) form channels facilitating the passive transport of water and other small polar molecules across membranes. In this study, the complete open reading frame (ORF) of CiMIP1 (GenBank ID KY316061) encoding one kind of MIPs in the Antarctic ice microalga Chlamydomonas sp. ICE-L is successfully cloned using RACE. In addition, the expression patterns of CiMIP1 gene under different conditions of temperature and salinity are determined by qRT-PCR. The ORF of CiMIP1 gene encodes 308 amino acids, and the deduced amino acid sequence shows 74% homology with Chlamydomonas reinhardtii CrMIP1 (GenBank number 159471952). Phylogenetic analysis reveals that algal MIPs are divided into seven groups, and it is speculated that CiMIP1 most likely belongs to the MIPD subfamily. In addition, we are surprised to find that a third NPA motif exists at the carboxy terminus of the target protein except for two highly conserved ones. Expression analysis shows that the transcriptional levels of CiMIP1 gene are upregulated under either lower temperature or higher temperature and high salinity. In summary, the results together have provide new insights into the newly discovered gene in green algae and lay the foundation for further studies on the adaptation mechanism of Chlamydomonas sp. ICE-L to abiotic stresses.

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Acknowledgements

This work was financially supported by the Natural Science Foundation of China (41576187), Basic Scientific Fund for National Public Research Institutes of China (2016Q10), Key Research and Development Program of Shandong Province (2016ZDJS06A03), Public Science and Technology Research Funds Projects of Ocean (201405015), Science and Technology Planning Project of Shandong Province (2014GHY115003), Major Projects of Independent Innovation Achievements Transformation in Shandong Province (2014ZZCX06202), Qingdao National Laboratory for Marine Science and Technology (2015ASKJ02), and Qingdao Entrepreneurship and Innovation Pioneers Program (15-10-3-15-(44)-zch).

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  1. Key Laboratory of Marine Bioactive Substances, First Institute of Oceanography, State Oceanic Administration, Xianxialing Road 6th, Qingdao, 266061, China

    Lulu Li, Changfeng Qu, Zhou Zheng, Yibin Wang, Fangming Liu, Yingying He, Xiaodong He & Jinlai Miao

  2. Medical College, Qingdao University, Ningxia Road 308th, Qingdao, 266071, China

    Meiling An & Jinlai Miao

  3. Laboratory for Marine Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China

    Jinlai Miao

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  1. Lulu Li
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  2. Meiling An
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Li, L., An, M., Qu, C. et al. Molecular cloning and expression analysis of major intrinsic protein gene in Chlamydomonas sp. ICE-L from Antarctica. Extremophiles 21, 817–827 (2017). https://doi.org/10.1007/s00792-017-0945-z

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