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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 31, No. 3, 2010, pp. 587-594
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Bioline Code: zr10081
Full paper language: Chinese
Document type: Research Article
Document available free of charge
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Zoological Research, Vol. 31, No. 3, 2010, pp. 587-594
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Cloning and expression pattern analysis of a lipopolysaccharide -and β-1,3-glucan-binding protein in Portunus trituberculatus
Ying, Zhang; Shan, Jin; Qing-Song, Zhao; Guo-Liang, Wang; Kai, Yu & Chun-Lin, Wang
Abstract
The lipopolysaccharide -and β-1,3-glucan-binding protein (LGBP) is a pattern recognition receptor, which is fundamental for the innate immune response of crustaceans. A LGBP gene was cloned from the haemocytes of Portunus trituberculatus using SMART RACE methods. The full-length LGBP cDNA (1 378 bp) had a 1 095 bp open reading frame encoding a protein of 365 amino acid residues including a 16 amino acid residues signal peptide, a 138 bp 5' untranslated region (UTR) and a 144 bp untranslated region in the 3' UTR with a 29 bp polyA tail. The calculated molecular mass of the mature protein (349 amino acid residues) is 39,825.24 with an estimated pI of 4.49. The gene sequence and secondary structure of LGBP were analyzed by bio-informatics. Additionally, a Glyco hydro 16 domain was identified. The expression of P. trituberculatus in various tissues were detected through RT-PCR methods. The results showed that the LGBP gene expressed in all the tissues detected, including haemocytes, hepatopancreas, heart, gills and muscle. In response to the challenge of Staphyloccocus aureus and Vibrio alginolyticus, the LGBP gene expression in haemocytes of the group challenged with mixed bacteria were higher than the control group within 48 h. It suggested that the LGBP gene plays an active role in immunologic process againsting bacterial infection.
Keywords
Portunus trituberculatus; LGBP; SMART RACE; RT-PCR
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