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Reference genes for qPCR expression in black tiger shrimp, Penaeus monodon

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Abstract

Background

Gene expression profiling via qPCR is an essential tool for unraveling the intricate molecular mechanisms underlying growth and development. Identifying and validating the most appropriate reference genes is essential for qPCR experiments. Nevertheless, there exists a deficiency in a thorough assessment of reference genes concerning the expression of the genes in the research in the context of the growth and development of the Black Tiger Shrimp, P. monodon. This popular marine crustacean is extensively raised for human consumption. In this study, we assessed the expression stability of seven reference genes (ACTB, 18S, EF-1α, AK, PK, cox1, and CLTC) in adult tissues (hepatopancreas, gills, and stomach) of small and large polymorphs of P. monodon.

Methods and results

The stability of gene expressions was assessed utilizing NormFinder, BestKeeper, and geNorm, and a comprehensive ranking of these genes was conducted through the online tool RefFinder. In the overall ranking, 18S and CLTC emerged as the most stable genes in the hepatopancreas and stomach, while CLTC and AK exhibited significant statistical reliability in the gills of adult P. monodon. The validation of these identified stable genes was carried out using a growth-associated gene, insr-1.

Conclusion

The results indicated that 18S and CLTC stand out as the most versatile reference genes for conducting qPCR analysis focused on the growth of P. monodon. This study represents the first comprehensive exploration that identifies and assesses reference genes for qPCR analysis in P. monodon, providing valuable tools for research involving similar crustaceans.

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Acknowledgements

The first author, Preety Sweta Hembrom acknowledges Rajiv Gandhi National Fellowship (RGNF) for providing fellowship. The Authors acknowledge research funding from the Central University of Kerala for this Project.

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  1. Department of Genomic Science, School of Biological Sciences, Central University of Kerala, Kasaragod, Kerala, 671316, India

    Preety Sweta Hembrom, Mottakunja Deepthi, Gourav Biswas, Bhagya Mappurath, Adon Babu, Narchikundil Reeja, Neeraja Mano & Tony Grace

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  1. Preety Sweta Hembrom
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Contributions

Conceptualization: T.G. and P.S.H.; Project administration: T.G.; Supervision: T.G.; Resources: P.S.H., N.M., and B.M.; Methodology: P.S.H., M.D., and A.B.; Formal Analysis: P.S.H and N.R.; Visualization: P.S.H and G.B.; Writing—original draft: P.S.H.; Writing—review & editing: T.G., M.D., and G.B. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Tony Grace.

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Hembrom, P.S., Deepthi, M., Biswas, G. et al. Reference genes for qPCR expression in black tiger shrimp, Penaeus monodon. Mol Biol Rep 51, 422 (2024). https://doi.org/10.1007/s11033-024-09409-5

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