Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper <italic style="font-style: italic">Cromileptes altivelis</italic>

Xiaojuan CHEN; Yun SUN; Panpan ZHANG; Jianlong LI; Haiping LI; Caoying WEI; Zhenjie CAO; Yongcan ZHOU

doi:10.1007/s00343-020-0238-8

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Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis

Biology | Updated：2023-04-27

- Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis
- Journal of Oceanology and Limnology Vol. 39, Issue 5, Pages: 1985-1999(2021)
- Affiliations：
  
  1.State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan University, Haikou 570228, China
  2.Key Laboratory of Tropical Hydrobiology and Biotechnology of Hainan Province, Haikou 570228, China
  3.Department of Aquaculture, College of Marine Sciences, Hainan University, Haikou 570228, China
- Author bio：
  
  Zhenjie CAO, E-mail: 1057319886@qq.com
  Yongcan ZHOU, E-mail: zychnu@163.com
- Funds：
- DOI：10.1007/s00343-020-0238-8
  CLC：
- Received：21 June 2020，
  
  Accepted：18 September 2020，
  
  Online First：13 November 2020，
  
  Published：2021-09
- Accepted：
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Xiaojuan CHEN, Yun SUN, Panpan ZHANG, et al. Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis[J]. Journal of Oceanology and Limnology, 2021, 39(5): 1985-1999.
DOI：

Xiaojuan CHEN, Yun SUN, Panpan ZHANG, et al. Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis[J]. Journal of Oceanology and Limnology, 2021, 39(5): 1985-1999. DOI： 10.1007/s00343-020-0238-8.

摘要

Abstract

Humpback grouper

Cromileptes altivelis

is one commercial fish with considerable economic value. To determine the expression stabilities of six commonly used internal reference genes in

altivelis

challenged by

Vibrio harveyi

and viral nervous necrosis virus (VNNV) through quantitative real-time PCR (qRT-PCR)

the expression levels of selected genes in five immune organs stimulated with pathogenic infection were carefully evaluated using algorithms of geNorm

NormFinder

and BestKeeper. The results show that the expression stabilities of the six candidate internal reference genes were different. Under normal physiological conditions

RPL13

were identified as the most stably expressed genes among five different immune organs (liver

spleen

kidney

intestine

and gill). After

harveyi

stimulation

RPL13

EF1A

RPL13

and

EF1A

were identified by geNorm

NormFinder

and BestKeeper as the most stable genes in liver

spleen

kidney

intestine

and gill

respectively. Combining these three algorithms suggested that under stimulation of

VNNV

RPL13

EF1A

Actin

RPL13

and

Actin

were as the most stable genes in liver

spleen

kidney

intestine

and gill

respectively. These results suggest that specific experiment conditions and tissue types shall be considered when selecting the reference genes in qRT-PCR analysis. This study provided a solid foundation for future studies on gene expression of

altivelis

under different conditions.

关键词

Keywords

references

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Related Institution

Hainan Academy of Ocean and Fisheries Sciences

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Screening of stable internal reference genes by quantitative real-time PCR in humpback grouper Cromileptes altivelis

DOI：10.1007/s00343-020-0238-8

摘要

Abstract

关键词

Keywords

references