Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata

Fei Pu; Bingye Yang; Caihuan Ke

doi:10.1007/s00343-015-4078-x

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Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata

Updated：2023-05-16

- Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata
- Journal of Oceanology and Limnology Vol. 33, Issue 4, Pages: 838-845(2015)
- Affiliations：
  
  1. State Key Laboratory of Marine Environmental Science Xiamen University
  2. College of Ocean and Earth Science Xiamen University
  3. Xiamen Medical College
- Author bio：
- Funds：
- DOI：10.1007/s00343-015-4078-x
  CLC：
- Online First：18 March 2015，
  
  Published：2015
- Accepted：
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Pu, F., Yang, B. & Ke, C. Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata, ., Chin. J. Ocean. Limnol. 33, 838–845 (2015). https://doi.org/10.1007/s00343-015-4078-x
DOI：

Pu, F., Yang, B. & Ke, C. Characterization of reference genes for qPCR analysis in various tissues of the Fujian oyster Crassostrea angulata, ., Chin. J. Ocean. Limnol. 33, 838–845 (2015). https://doi.org/10.1007/s00343-015-4078-x DOI：

摘要

Abstract

Accurate quantification of transcripts using quantitative real-time polymerase chain reaction (qPCR) depends on the identification of reliable reference genes for normalization. This study aimed to identify and validate seven reference genes

including actin-2 (ACT-2)

elongation factor 1 alpha (EF-1α)

elongation factor 1 beta (EF-1β)

glyceraldehyde-3-phosphate dehydrogenase (GAPDH)

ubiquitin (UBQ)

β-tubulin (β-TUB)

and 18S ribosomal RNA

from Crassostrea angulata

a valuable marine bivalve cultured worldwide. Transcript levels of the candidate reference genes were examined using qPCR analysis and showed differential expression patterns in the mantle

gill

adductor muscle

labial palp

visceral mass

hemolymph and gonad tissues. Quantitative data were analyzed using the geNorm software to assess the expression stability of the candidate reference genes

revealing that β-TUB and UBQ were the most stable genes. The commonly used GAPDH and 18S rRNA showed low stability

making them unsuitable candidates in this system. The expression pattern of the G protein β-subunit gene (Gβ) across tissue types was also examined and normalized to the expression of each or both of UBQ and β-TUB as internal controls. This revealed consistent trends with all three normalization approaches

thus validating the reliability of UBQ and β-TUB as optimal internal controls. The study provides the first validated reference genes for accurate data normalization in transcript profiling in Crassostrea angulata

which will be indispensable for further functional genomics studies in this economically valuable marine bivalve.

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references

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