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Molecular cloning and expression of a heat-shock cognate 70 (hsc70) gene from swordtail fish (Xiphophorus helleri)

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Abstract

Heat shock proteins are a family of molecular chaperones that are involved in many aspects of protein homeostasis. In the present study, a full-length cDNA, encoding the constitutively expressed 70-kDa heat shock cognate protein (Hsc70), was isolated from swordtail fish (Xiphophorus helleri) and designated as XheHsc70. The Xhehsc70 cDNA was 2 104 bp long with an open reading frame of 1 941 bp, and it encoded a protein of 646 amino acids with a theoretical molecular weight of 70.77 kDa and an isoelectric point of 5.04. The deduced amino acid sequence shared 94.1%-98.6% identities with the Hsc70s from a number of other fish species. Tissue distribution results show that the Xhehsc70 mRNA was expressed in brain, heart, head kidney, kidney, spleen, liver, muscle, gill, and peripheral blood. After immunization with formalin-killed Vibrio alginolyticus cells there was a significant increase in the Xhehsc70 mRNA transcriptional level in the head kidney of the vaccinated fish compared with in the control at 6, 12, 24, and 48 h as shown by quantitative real time RT-PCR. Based on an analysis of the amino acid sequence of XheHsc70, its phylogeny, and Xhehsc70 mRNA expression, XheHsc70 was identified as a member of the cytoplasmic Hsc70 (constitutive) subfamily of the Hsp70 family of heat shock proteins, suggesting that it may play a role in the immune response. The Xhehsc70 cDNA sequence reported in this study was submitted to GenBank under the accession number JF739182.

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Authors and Affiliations

  1. Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Fishery Drug Development, Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province, Guangzhou, 510380, China

    Ningqiu Li  (李宁求), Xiaozhe Fu  (付小哲), Jingang Han  (韩进刚), Cunbin Shi  (石存斌), Zhibin Huang  (黄志斌) & Shuqin Wu  (吴淑勤)

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  1. Ningqiu Li  (李宁求)
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  2. Xiaozhe Fu  (付小哲)
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  3. Jingang Han  (韩进刚)
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  4. Cunbin Shi  (石存斌)
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  5. Zhibin Huang  (黄志斌)
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  6. Shuqin Wu  (吴淑勤)
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Correspondence to Shuqin Wu  (吴淑勤).

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Supported by the National Key Technology R&D Program of China (No. 2012BAD25B02) and the Natural Science Foundation of Guangdong Province of China (Nos. 7004728, 06024033) LI Ningqiu and FU Xiaozhe contributed equally to this paper.

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Li, N., Fu, X., Han, J. et al. Molecular cloning and expression of a heat-shock cognate 70 (hsc70) gene from swordtail fish (Xiphophorus helleri). Chin. J. Ocean. Limnol. 31, 821–829 (2013). https://doi.org/10.1007/s00343-013-2208-x

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