Sequence Localization of SeGPx in S. digitata Genome Contigs and Determination of its Presence in the Whole Worm Extract
- PMID: 40694172
- DOI: 10.1007/s11686-025-01104-0
Sequence Localization of SeGPx in S. digitata Genome Contigs and Determination of its Presence in the Whole Worm Extract
Abstract
Purpose: Lymphatic filariasis, caused by three major filarial species, is marked by immune evasion strategies involving antioxidant enzymes. The role of selenium-dependent glutathione peroxidase (SeGPx) in this process remains underexplored. This study aimed to identify and characterise SeGPx in Setaria digitata, a genomic analogue of Wuchereria bancrofti, and evaluate its potential as a diagnostic antigen.
Methods: segpx sequences were identified through bioinformatics tools, including BLAST and UniProt databases. Due to limited nematode entries, a validated SeGPx sequence from Lymnaea stagnalis was used as a proxy in PSI-BLAST to identify homologues. Enzymatic activity was confirmed through spectrophotometric assay and activity staining on SDS-PAGE to confirm its enzymatic activity and molecular mass confirmation.
Results: The segpx gene was localised within the S. digitata genome data (Contig 127, nucleotides 56,000-58,000). The enzyme assay showed a time-dependent decline in absorbance at 340 nm due to NADPH oxidation, plateauing after 13 min. Enzyme activity was calculated as 0.139 U, with a specific activity of 0.198 U/mg protein. A clear band at ~ 20 kDa was visualised via activity staining, confirming SeGPx presence.
Conclusion: Combined sequence-based identification and enzymatic validation confirm the functional presence of SeGPx in S. digitata. These findings support its role in oxidative stress mitigation and potential as a diagnostic antigen. The precise gene localisation offers a foundation for recombinant cloning, providing a streamlined alternative to conventional purification approaches for diagnostic development in filariasis.
Keywords: Setaria digitata; Diagnostic antigen; Lymphatic filariasis; Selenium glutathione peroxidase.
© 2025. The Author(s), under exclusive licence to Springer Nature Switzerland AG.
Conflict of interest statement
Declarations. Competing Interests: The authors declare no competing interests.
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