闽江学院学报2024,Vol.45Issue(5) :67-79.DOI:10.19724/j.cnki.jmju.2024.05.008

不同株系球形棕囊藻qRT-PCR内参基因的筛选

Screening Suitable Reference Genes for qRT-PCR in Different Strains of Phaeocystis globosa

许歆 王锦荣 许欣红 徐肖 李靖 杨杰
闽江学院学报2024,Vol.45Issue(5) :67-79.DOI:10.19724/j.cnki.jmju.2024.05.008

不同株系球形棕囊藻qRT-PCR内参基因的筛选

Screening Suitable Reference Genes for qRT-PCR in Different Strains of Phaeocystis globosa

许歆 1王锦荣 2许欣红 1徐肖 1李靖 1杨杰1
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作者信息

  • 1. 闽江学院地理与海洋学院,福建省海洋生物多样性保护与永续利用重点实验室,福建 福州 350108
  • 2. 中国冶金地质总局第二地质勘查院,福建 福州 350108
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摘要

研究了 7 个候选内参基因(α-tubulin、β-tubulin、Actin、Ubiquitin、18S rRNA、EF-1α和GAPDH),在3 株成囊能力不同的球形棕囊藻(成囊株系PG-1 和PG-2 以及不成囊的单细胞株系PG-3)不同生长阶段下的表达稳定性.结果表明:PG-1 的最佳内参基因组合为Ubiquitin、β-tubulin和18S rRNA;PG-2 的最佳内参基因组合为α-tubulin、18S rRNA、β-tubulin和EF-1α,可见在成囊株系中表达较为稳定的内参基因为β-tubulin和 18S rRNA;在单细胞株系PG-3 中表达最稳定的内参基因为Ubiquitin和α-tubulin;对3 个株系球形棕囊藻的基因表达差异进行标准化分析可使用EF-1α、Ubiquitin和 18S rRNA为内参基因组合.所得结论为更准确地研究不同株系球形棕囊藻在不同生长时期的基因表达、验证成囊的关键基因奠定基础.

Abstract

Expression stability of seven candidate reference genes in different growth stages of three Pha ̄eocystis globosa strains with different colony-formation ability was studied.Results showed that the optimal reference genes in colony-forming strain PG-1 are Ubiquitin,β-tubulin and 18S rRNA.α-tubulin,18S rRNA,β-tubulin and EF-1α are the best combination for another colony-forming strain PG-2.Therefore,β-tubulin and 18S rRNA were stably expressed in the colony-forming strains.The most stable reference genes in non-colony-forming strain PG-3 were Ubiquitin and α-tubulin.EF-1α,Ubiquitin and 18S rRNA could be used for the standardized analysis of gene expression level among three P.globosa strains.This study provides a basis for a more accurate study on the gene expression levels of P.globosa in different strains and different growth stages,especially verifying the key genes of colony formation.

关键词

球形棕囊藻/不同株系/内参基因/实时荧光定量PCR

Key words

Phaeocystis globosa/different strains/reference genes/qRT-PCR

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出版年

2024
闽江学院学报
闽江学院

闽江学院学报

CHSSCD
影响因子:0.221
ISSN:1009-7821
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