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Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction


Citation

Sim, Mei Chea and Cheong, Xin Chan and Ho, Chai Ling and Phang, Siew Moi (2018) Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction. Phycological Research, 66 (4). 247 - 252. ISSN 1322-0829; ESSN: 1440-1835

Abstract

Sargassum species are one of the major alginate-producing seaweed species in Asian countries. Alginate is widely used in food, feed, pharmaceutical and medical industries as thickening and stabilizing agents. To establish a set of consistently expressed genes as reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) studies of Sargassum polycystum (Fucales, Ochrophyta) in samples collected at two distinct time points from the field, four candidate reference genes, namely ribosomal protein L3 (RPL3), ribosomal protein S15 (RPS15), alphatubulin (α-TUB) and eukaryotic translation elongation factor 1 alpha (TEF1α), were analyzed using geNorm and NormFinder. The results showed that RPL3, α-TUB and TEF1α were the most stable genes using both programs, whereas RPS15 gene was shown to be the least stable. Identification of stably expressed reference genes is crucial for qRT-PCR studies to allow accurate quantification of target gene expression levels. In addition, the expression of key enzyme in the final step of alginate biosynthesis pathway mannuronan C5 epimerase-SP01411 (MC5E-SP01411) and mannuronan C5 epimerase-SP02271 (MC5E-SP02271) were differentially expressed in the seaweeds collected at two distinct time points from the field. To our knowledge, this is the first report on validation of reference genes for any Sargassum species. Our data provide a basis for the selection of reference genes for future biological research in related studies.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
DOI Number: https://doi.org/10.1111/pre.12328
Publisher: Wiley
Keywords: Brown algae; Mannuronan C5-epimerase (MC5E); Normalization; qRT-PCR; Reference genes
Depositing User: Ms. Nida Hidayati Ghazali
Date Deposited: 15 Jul 2021 23:05
Last Modified: 15 Jul 2021 23:05
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1111/pre.12328
URI: http://psasir.upm.edu.my/id/eprint/73851
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