Cloning, Expression and Stability Analysis of Candidate Reference Genes in the Glacial Relict Hucho bleekeri Under Immune Stress

IF 0.7 4区农林科学 Q4 FISHERIES

Journal of Applied Ichthyology Pub Date : 2025-07-22 DOI:10.1155/jai/5526431

Xiaoyun Wu, Jiansheng Lai, Yeyu Chen, Huanchao Yang, Yanling Chen, Zhao Liu, Quanyu Tu, Yi Yu, Hua Li

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Abstract

To find reference genes (RGs) and establish a screening method for RGs in Sichuan taimen (Hucho bleekeri), six candidate RGs—β-actin, b2m, β-tubulin, ef1α, gapdh and hprt — were cloned, and the stability of expression of the six candidate RGs were analysed. The six RG sequences in H. Bleekeri had higher similarity with the corresponding sequences in Salmo or Oncorhynchus. The RGs geomean of ranking values of gene expression in tissues of H. bleekeri was ef1α > β-actin > β-tubulin > hprt > b2m > gapdh. After lipopolysaccharide (LPS) treatment, the RGs geomean of ranking values of gene expression in spleen was ef1α > β-actin > b2m > gapdh > β-tubulin > hprt, in head kidney was β-actin > ef1α > β-tubulin > hprt > gapdh > b2m, in liver was β-tubulin > hprt > β-actin > gapdh > ef1α > b2m; after polyinosinic: polycytidylic acid (Poly (I:C)) treatment, the values in spleen was ef1α > hprt > β-actin > b2m > β-tubulin > gapdh, in head kidney was β-actin > ef1α > β-tubulin > hprt > gapdh > b2m, in liver was β-tubulin > β-actin > ef1α > gapdh > b2m > hprt. The results indicated that ef1α can be used as the most stable RG for the tissue distribution analysis of gene expression in H. bleekeri, and ef1α, β-actin and β-tubulin was suitable for spleen, head kidney and liver cells, respectively, under LPS and Ploy (I: C) stimulation.

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免疫胁迫下冰川残体Hucho bleekeri候选内参基因的克隆、表达及稳定性分析

为了寻找四川台菜（Hucho bleekeri）的内参基因（RGs），建立四川台菜（Hucho bleekeri） RGs的筛选方法，克隆了6个候选RGs - β-actin、b2m、β-tubulin、ef1α、gapdh和hprt -，并分析了6个候选RGs的表达稳定性。Bleekeri的6个RG序列与Salmo和Oncorhynchus的相似度较高。白僵菌组织中基因表达排序值的RGs几何为ef1α >； β-肌动蛋白>； β-微管蛋白>； hprt > b2m > gapdh。脂多糖（LPS）处理后，脾脏基因表达的RGs排序值为ef1α >； β-肌动蛋白>； b2m > gapdh >； β-微管蛋白>； hprt；头肾为β-肌动蛋白>； ef1α > hprt; gapdh > b2m，肝脏为β-微管蛋白>； hprt >； β-肌动蛋白>； gapdh > b2m；多肌苷：多胞酸（Poly (I:C)）处理后，脾脏值为ef1α >； hprt >； β-肌动蛋白>； b2m >； β-微管蛋白>； gapdh > gapdh; b2m；肝组织值为β-微管蛋白>； β-肌动蛋白>； ef1α > gapdh > b2m > hprt。结果表明，在LPS和poly （I: C）刺激下，ef1α、β-肌动蛋白和β-微管蛋白分别适用于脾、头肾和肝细胞，ef1α可作为最稳定的RG进行基因表达的组织分布分析。

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来源期刊

Journal of Applied Ichthyology 生物-海洋与淡水生物学

CiteScore

2.30

自引率

11.10%

发文量

审稿时长

3-8 weeks

期刊介绍： The Journal of Applied Ichthyology publishes articles of international repute on ichthyology, aquaculture, and marine fisheries; ichthyopathology and ichthyoimmunology; environmental toxicology using fishes as test organisms; basic research on fishery management; and aspects of integrated coastal zone management in relation to fisheries and aquaculture. Emphasis is placed on the application of scientific research findings, while special consideration is given to ichthyological problems occurring in developing countries. Article formats include original articles, review articles, short communications and technical reports.