Characterization of the BPI-like gene from a subtracted cDNA library of large yellow croaker (Pseudosciaena crocea

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• 作者：Yanqing Huang ; Huifang Lou ; Xinzhong Wu ; Yanxia Chen
• 关键词：Large yellow croaker (Pseudosciaena crocea) ; Suppression subtraction hybridization ; Bactericidal permeability-increasing protein ; Lipopolysaccharide ; Pc-BPI-like gene ; Eukaryotic expression
• 刊名：Fish & Shellfish Immunology
• 出版年：2008
• 出版时间：December 2008
• 年：2008
• 卷：25
• 期：6
• 页码：740-750
• 全文大小：1204 K

文摘

One expressed sequence tag (EST 64LF004 clone), which is from the subtracted cDNA library of the head kidney of large yellow croaker (Pseudosciaena crocea) stimulated with peptidoglycan (PG) by suppression subtractive hybridization (SSH) method, was cloned using RACE–PCR. The full length cDNA, which possesses typical structural features of a signal peptide, a conserved LPS binding domain and two bactericidal permeability-increasing (BPI) motifs as in higher vertebrates, was identified as a novel homologue, namely of the large yellow croaker BPI-like molecule (Pc-BPI-L). Phylogenetic analysis showed this Pc-BPI-L of large yellow croaker as the most ancestral branch in bony fish clade. The recombinant Pc-BPI-L protein expressed in the Tn-5B1-4 insect cells was successfully produced and confirmed to have the predicted size of 52 kDa by Western blot analysis. At the message level, Pc-BPI-L mRNA was ubiquitously expressed in all tissues examined. Following formalin-inactivated Vibrio alginolyticus and Nocardia seriolae treatment, Pc-BPI-L message was differentially up-regulated in primary immune organs. These results indicate that Pc-BPI-L might be involved in the immune response to bacterial infection.