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

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...

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Main Authors: Sim, Mei Chea, Cheong, Xin Chan, Ho, Chai Ling, Phang, Siew Moi
Format: Article
Language:English
Published: Wiley 2018
Online Access:http://psasir.upm.edu.my/id/eprint/73851/
http://psasir.upm.edu.my/id/eprint/73851/1/ALGAE.pdf
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author Sim, Mei Chea
Cheong, Xin Chan
Ho, Chai Ling
Phang, Siew Moi
author_facet Sim, Mei Chea
Cheong, Xin Chan
Ho, Chai Ling
Phang, Siew Moi
author_sort Sim, Mei Chea
building UPM Institutional Repository
collection Online Access
description 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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spelling upm-738512021-07-15T23:05:36Z http://psasir.upm.edu.my/id/eprint/73851/ Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction Sim, Mei Chea Cheong, Xin Chan Ho, Chai Ling Phang, Siew Moi 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. Wiley 2018-10 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/73851/1/ALGAE.pdf 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 https://onlinelibrary.wiley.com/doi/abs/10.1111/pre.12328 10.1111/pre.12328
spellingShingle Sim, Mei Chea
Cheong, Xin Chan
Ho, Chai Ling
Phang, Siew Moi
Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title_full Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title_fullStr Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title_full_unstemmed Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title_short Selection of reference genes for transcript profiling of Sargassum polycystum by quantitative real-time polymerase chain reaction
title_sort selection of reference genes for transcript profiling of sargassum polycystum by quantitative real-time polymerase chain reaction
url http://psasir.upm.edu.my/id/eprint/73851/
http://psasir.upm.edu.my/id/eprint/73851/
http://psasir.upm.edu.my/id/eprint/73851/
http://psasir.upm.edu.my/id/eprint/73851/1/ALGAE.pdf