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Molecular characterization and expression analysis of lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) from pearl oyster Pinctada fucata

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Abstract

The lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) plays an important function in the innate immune response of invertebrates as a pattern recognition receptor (PRR). Herein, we described the isolation and characterization of pearl oyster Pinctada fucata LGBP (designated as poLGBP). The poLGBP cDNA was 2,075 bp long and consisted of a 5′-untranslated region (UTR) of 18 bp, a 3′-UTR of 299 bp with one cytokine RNA instability motifs (ATTTA), and an open reading frame (ORF) of 1,758 bp encoding a polypeptide of 585 amino acids with an estimated molecular mass of 65.1 kDa and a theoretical isoelectric point of 5.80. Homology analysis of the deduced amino acid sequence of the poLGBP with other known LGBP sequences by MatGAT software revealed that the poLGBP shared 26.3–56.7% identity and 40.5–70.9% similarity to the other known LGBP sequences. SMART and alignment analysis revealed that the poLGBP possessed a potential polysaccharide-binding motif, a glucanase motif, a LPS-binding site, a β-1,3-linkage of polysaccharide, a glycine-rich region, a threonine-rich region and two N-glycosylation sites. In healthy pearl oyster, the poLGBP mRNA was specifically expressed in digestive gland, and not detected in gill, adductor muscle, gonad, intestine, mantle and hemocytes. However, after bacteria stimulation, the expression of the poLGBP mRNA was significantly up-regulated in digestive gland and also weakly detected in haemocytes, gonad and intestine. After LPS stimulation, the poLGBP mRNA expression was significantly up-regulated at 8 and 12 h in digestive gland, and the expression level was 10.7-fold higher than the PBS group at 12 h. After bacteria stimulation, the expression level of the poLGBP mRNA was also significantly up-regulated in digestive gland and was 12.9-fold higher than the PBS group at 8 h. However, during the experiment, the poLGBP mRNA expression was not detected in gill after LPS or bacteria stimulation. The tissue-specific expression and the expression up-regulation after LPS or bacteria stimulation in digestive gland suggested that the poLGBP was an inducible acute-phase protein and might play an important function in digestion as digestive enzyme and pattern recognition receptor.

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Acknowledgments

We are grateful to all the laboratory members for their technical advice and helpful discussions. This research was supported by the major science and technology projects of Guangdong (A200701C02; 2008A020100004), national sci-tech platform projects (2005DKA30470) and central institutes of public welfare projects (ZD-02; 2009TS23).

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Authors and Affiliations

  1. Division of Aquaculture and Biotechnology, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Science, 510300, Guangzhou, China

    Dianchang Zhang, Jianjun Ma, Jingjing Jiang, Lihua Qiu, Caiyan Zhu, Tianfeng Su, Youning Li, Kaichang Wu & Shigui Jiang

  2. School of Life Science and Technology, Jinan University, 510632, Guangzhou, China

    Jianjun Ma

  3. School of Fisheries and Life Science, Shanghai Ocean University, 200090, Shanghai, China

    Jingjing Jiang

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  1. Dianchang Zhang
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Correspondence to Shigui Jiang.

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Dianchang Zhang and Jianjun Ma authors contributed equally to this work.

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Zhang, D., Ma, J., Jiang, J. et al. Molecular characterization and expression analysis of lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) from pearl oyster Pinctada fucata . Mol Biol Rep 37, 3335–3343 (2010). https://doi.org/10.1007/s11033-009-9920-9

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