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<p>Grouper species are some of the most valuable aquaculture fish worldwide. However, their vulnerability to different pathogens causes considerable economic losses. The galectin-9 gene was identified in the transcriptome of the seven-band grouper <i>Hyporthodus septemfasciatus</i>. The galectin-9 gene was cloned, and its recombinant protein was overexpressed, enabling a detailed analysis of its molecular characteristics. The complete cDNA sequence consists of 2,005 base pairs (bp), encompassing a 162&#xa0;bp 5′-untranslated region (UTR), an 829&#xa0;bp 3′-UTR, and a 1,014&#xa0;bp open reading frame (ORF) that encodes 338 amino acids (aa). SGGal-9 possesses two conserved carbohydrate recognition domains (CRDs): an N-terminal CRD comprising 135 aa and a C-terminal CRD comprising 123 aa. Each CRD in SGGal-9 features two conserved β-galactoside binding motifs (H_NPR and WG_EER). The predicted protein structure lacked signal peptides or transmembrane domains. After sub-cloning the SGGal-9 gene into the pCold I vector, recombinant SGGal-9 (rSGGal-9) was successfully over-expressed as a soluble protein in <i>Escherichia coli (E. coli)</i> BL21 (DE3), induced with 0.1&#xa0;mM isopropyl-β-D-1-thiogalactopyranoside (IPTG). To assess rSGGal-9 activity, hemagglutination assays were employed using erythrocytes from olive flounder (<i>Paralichthys olivaceus</i>) and sevenband grouper and demonstrated effective aggregation at minimum concentrations of 3.125&#xa0;μg/mL and 12.5&#xa0;μg/mL, respectively. Furthermore, the aggregation capability of rSGGal-9 was confirmed in different bacteria, including <i>E. coli, Lactococcus garvieae</i>, <i>Photobacterium damselae</i>, <i>Streptococcus iniae</i>, <i>Streptococcus parauberis</i>, and <i>Vibrio parahaemolyticus</i>. These findings, based solely on in vitro assays, suggest that galectin-9 has bacterial-aggregating properties; however, further in vivo studies are required to evaluate its preventive potential in aquaculture.</p>

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Molecular characterization and in vitro bacterial aggregating properties of tandem-repeat galectin-9 gene from sevenband grouper (Hyporthodus septemfasciatus)

  • 期刊论文

  • 发表日期：

    2026年1月27日

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  • Kyungmin Kang,
  • Jeongmin Seo,
  • Maheshkumar Prakash Patil,
  • Seung Hyun Yoo,
  • Myung-Joo Oh,
  • Jong-Oh Kim

摘要

Grouper species are some of the most valuable aquaculture fish worldwide. However, their vulnerability to different pathogens causes considerable economic losses. The galectin-9 gene was identified in the transcriptome of the seven-band grouper Hyporthodus septemfasciatus. The galectin-9 gene was cloned, and its recombinant protein was overexpressed, enabling a detailed analysis of its molecular characteristics. The complete cDNA sequence consists of 2,005 base pairs (bp), encompassing a 162 bp 5′-untranslated region (UTR), an 829 bp 3′-UTR, and a 1,014 bp open reading frame (ORF) that encodes 338 amino acids (aa). SGGal-9 possesses two conserved carbohydrate recognition domains (CRDs): an N-terminal CRD comprising 135 aa and a C-terminal CRD comprising 123 aa. Each CRD in SGGal-9 features two conserved β-galactoside binding motifs (H_NPR and WG_EER). The predicted protein structure lacked signal peptides or transmembrane domains. After sub-cloning the SGGal-9 gene into the pCold I vector, recombinant SGGal-9 (rSGGal-9) was successfully over-expressed as a soluble protein in Escherichia coli (E. coli) BL21 (DE3), induced with 0.1 mM isopropyl-β-D-1-thiogalactopyranoside (IPTG). To assess rSGGal-9 activity, hemagglutination assays were employed using erythrocytes from olive flounder (Paralichthys olivaceus) and sevenband grouper and demonstrated effective aggregation at minimum concentrations of 3.125 μg/mL and 12.5 μg/mL, respectively. Furthermore, the aggregation capability of rSGGal-9 was confirmed in different bacteria, including E. coli, Lactococcus garvieae, Photobacterium damselae, Streptococcus iniae, Streptococcus parauberis, and Vibrio parahaemolyticus. These findings, based solely on in vitro assays, suggest that galectin-9 has bacterial-aggregating properties; however, further in vivo studies are required to evaluate its preventive potential in aquaculture.