Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers <italic style="font-style: italic">Plectropomus leopardus</italic>

Han ZHANG; Haoran YANG; Pengsuo LI; Na ZHAO; Xiaojuan CHEN; Yongcan ZHOU; Zhenjie CAO; Yun SUN

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Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers Plectropomus leopardus

Biology | Updated：2024-10-12

- Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers Plectropomus leopardus
- Journal of Oceanology and Limnology Vol. 41, Issue 3, Pages: 1084-1099(2023)
- 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：
  
  1057319886@qq.com
  syshui207@126.com
- Funds：
- DOI：
  CLC：
- Received：20 January 2022，
  
  Published：01 May 2023
- Accepted：
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ZHANG Han,YANG Haoran,LI Pengsuo,et al.Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers ,Plectropomus leopardus[J].Journal of Oceanology and Limnology,2023,41(03):1084-1099.
DOI：

ZHANG Han,YANG Haoran,LI Pengsuo,et al.Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers ,Plectropomus leopardus[J].Journal of Oceanology and Limnology,2023,41(03):1084-1099. DOI：

摘要

Abstract

Leopard coral groupers (

Plectropomus leopardus

)

commercially bred in South China

are a significant economical fish species. In this study

by means of quantitative real-time PCR (qRT-PCR) technology

we assessed the stability of six common reference genes expression and selected the appropriate reference genes in leopard coral groupers with or without

Vibrio harveyi

stimulation at different time points. These data produced by qRT-PCR was handled via the geNorm

NormFinder

and BestKeeper software. The results revealed all the examined reference genes had manifest tissue-specific expression in different tissues. Prior to

harveyi

stimulat

ion

RPL13

gene was the appropriate reference gene among eleven tissue types (blood

spleen

hepatopancreas

kidney

stomach

gill

heart

skin

muscle

intestine

brain) in leopard coral groupers. Under

harveyi

stimulation

the most reliable reference genes varied from tissue to tissue and were closely hinged upon different time points. At 6-h post-bacterial injection

the appropriate reference genes in hepatopancreas

spleen

kidney

and gill were

Actin

B2M

UBCE

and

Actin

respectively. At 9- and 12-h post-bacterial injection

the appropriate reference genes in hepatopancreas

spleen

kidney

and gill were

RPL13

Actin

and

Actin

respectively. If one reference gene was preferable

RPL13

Actin

and

Actin

could be selected as the reference gene in hepatopancreas

spleen

kidney

and gill of leopard coral groupers after

harveyi

infection

respectively. Expression profiles of two target genes (

and

lysin

) were used to further validate reliability of these selected appropriate candidates. This study will lay a solid foundation for the future research on qRT-PCR analysis of gene expression in leopard coral groupers.

关键词

Keywords

references

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⁰

Selection of the appropriate reference genes by quantitative real-time PCR in leopard coral groupers Plectropomus leopardus

DOI：

摘要

Abstract

关键词

Keywords

references