Identification of suitable reference genes for quantitative gene expression analysis in clam <italic style="font-style: italic">Cyclina sinensis</italic> under salinity stress and <italic style="font-style: italic">Vibrio</italic> infection

Fengjuan JIANG; Qingyao WANG; Jingjing DU; Fu LÜ; Qing NIE; Weihong ZHAO

doi:10.1007/s00343-021-1335-z

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Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection

Aquaculture and Fisheries | Updated：2023-04-27

- Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection
- Journal of Oceanology and Limnology Vol. 41, Issue 1, Pages: 352-363(2023)
- Affiliations：
  
  College of Marine and Biological Engineering, Yancheng Institute of Technology, Yancheng 224051, China
- Author bio：
  
  Qing NIE, E-mail: nieqing0422@163.com
  Weihong ZHAO, E-mail: misszwh@163.com
- Funds：
- DOI：10.1007/s00343-021-1335-z
  CLC：
- Received：13 October 2021，
  
  Accepted：07 December 2021，
  
  Online First：21 December 2021，
  
  Published：2023-01
- Accepted：
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Fengjuan JIANG, Qingyao WANG, Jingjing DU, et al. Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection[J]. Journal of Oceanology and Limnology, 2023, 41(1): 352-363.
DOI：

Fengjuan JIANG, Qingyao WANG, Jingjing DU, et al. Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection[J]. Journal of Oceanology and Limnology, 2023, 41(1): 352-363. DOI： 10.1007/s00343-021-1335-z.

摘要

Abstract

The appropriate reference gene is a prerequisite for accurate normalization of gene expression level

and research on suitable reference genes in clam

Cyclina sinensis

is scarce. To improve the situation

we selected five commonly used housekeeping genes

including

actin

Elongation factor 1-

(

EF1

)

Glyceraldehyde-3-phosphate dehydrogenase (

GAPDH

)

40S ribosomal protein S18 (

RPS18

)

and Tubulin

(

TUB

)

then evaluated their expression stability in different adult tissues and under different experimental treatments (salinity stress and

Vibrio

parahaemolyticus

infection). Their expression stability was analyzed by three frequently used programs

geNorm

NormFinder

and BestKeeper. This analysis indicated that multiple genes should be used for normalization

and we concluded that the reference gene combination

GAPDH

RPS18

actin

should be used for qRT-PCR analysis in different tissues of

sinensis

under normal physiological conditions. For the clams under salinity stress and

Vibrio

infection

EF1

GAPDH

RPS18

was recommended as the gene combination for qRT-PCR normalization.

TUB

was generally poorly ranked by all programs

and should not be used in future studies. This study should provide fundamental support for accurate quantitative gene expression analysis of this species.

关键词

Keywords

references

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Identification of suitable reference genes for quantitative gene expression analysis in clam Cyclina sinensis under salinity stress and Vibrio infection

DOI：10.1007/s00343-021-1335-z

摘要

Abstract

关键词

Keywords

references