Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano <italic style="font-style: italic">Trachinotus blochii</italic>

Xiaojuan CHEN; Xiaoqi ZHANG; Yun SUN; Zhigang TU; Zhenjie CAO; Shifeng WANG; Yongcan ZHOU

doi:10.1007/s00343-019-9053-5

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Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

Updated：2023-04-27

- Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii
- Journal of Oceanology and Limnology Vol. 38, Issue 2, Pages: 480-489(2020)
- Affiliations：
  
  1.State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, China
  2.Hainan Academy of Ocean and Fisheries Sciences, Haikou 570228, China
- Author bio：
  
  SUN Yun, syshui207@126.com
  ZHOU Yongcan, zychnu@163.com
- Funds：
- DOI：10.1007/s00343-019-9053-5
  CLC：
- Received：04 March 2019，
  
  Accepted：24 May 2019，
  
  Online First：05 August 2020，
  
  Published：2020-03
- Accepted：
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Xiaojuan CHEN, Xiaoqi ZHANG, Yun SUN, et al. Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii[J]. Journal of Oceanology and Limnology, 2020, 38(2): 480-489.
DOI：

Xiaojuan CHEN, Xiaoqi ZHANG, Yun SUN, et al. Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii[J]. Journal of Oceanology and Limnology, 2020, 38(2): 480-489. DOI： 10.1007/s00343-019-9053-5.

摘要

Abstract

Trachinotus blochii

is one of the important commercial fish species. In this study

we aim to confirm the reliability reference genes in

T. blochii

during different bacterial challenge through quantitative real-time PCR (qRT-PCR). The expression of the seven selected genes in four immune organs (i.e.

spleen

kidney

intestine

and gill) stimulated with

Vibrio harveyi

Edwardsiella tarda

and

Streptococcus agalactiae

were determined by qRT-PCR. The PCR data was analyzed using the geNorm and NormFinder algorithms. The results showed the selection of the internal controls should be tissue specific when studying gene expression in response to bacterial stimulation. After 48 h of stimulation with

V. harveyi

geNorm ranked EF1A/Actin

18S rRNA/B2M

UBCE/B2M

and 18S rRNA/B2M

as the most stably expressed genes in spleen

kidney

intestine

and gill

respectively. After 48 h of stimulation with

E. tarda

geNorm ranked 18S rRNA/EF1A

18S rRNA/B2M

B2M/RPL13

and 18S rRNA/EF1A

as the most stably expressed genes in spleen

kidney

intestine

and gill

respectively. After 48 h of stimulation with

S. agalactiae

18S rRNA/ EF1A

18S rRNA/B2M

B2M/Actin

and 18S rRNA/B2M were ranked as the most stably expressed genes in spleen

kidney

intestine

and gill

respectively. Compared to the results analyzed by geNorm

reference genes received similar rankings when using NormFinder software. The results showed that the reference genes appeared to be not only tissue specific

but also specific to the infecting species of bacteria. If one gene is preferred when

T. blochii

were infected by bacteria

18S rRNA

B2M

18S rRNA may be used in spleen

kidney

intestine

and gill

respectively.

关键词

Keywords

references

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Related Institution

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Assessment of internal controls for data normalization of gene expression after different bacterial stimulation by quantitative real-time PCR in golden pompano Trachinotus blochii

DOI：10.1007/s00343-019-9053-5

摘要

Abstract

关键词

Keywords

references