Selection of reference genes for quantitative real-time PCR in Octopus minor (Cephalopoda: Octopoda) under acute ammonia stress
Selection of reference genes for quantitative real-time PCR in Octopus minor (Cephalopoda: Octopoda) under acute ammonia stress
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急性氨胁迫下小章鱼（头足类：章鱼）实时定量 PCR 内参基因的选择
DOI：
10.1016/j.etap.2018.04.010
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发表时间：
2018
影响因子：
4.3
通讯作者：
Zheng Xiaodong
中科院分区：
文献类型：
--
作者：
Xu Ran;Zheng Xiaodong
高浓度氨氮是水产养殖业普遍存在的问题，往往对水产品造成不利影响。探索急性氨应激相关基因的表达调控有助于理解氨反应的分子机制。带有合适参考基因的实时定量聚合酶链式反应(qRT-PCR)是实现靶基因表达正常化的有效方法。为了确定小章鱼(软体动物：头足纲：章鱼)在急性氨胁迫和正常养殖条件下最适合的参考基因，选择了9个候选基因进行验证：OD、RPS18b、RPL29、RPS5、EF-1、α、RPL6、AA4、ACT、TUB。结果表明，候选基因在鳃、消化腺、脑和血淋巴中的稳定性差异较大。RPS5和RPL6在鳃中表现出较高的稳定性，RPL6、TUB、RPL29和OD是消化腺中四个合适的参考基因，EF-1α、TUB和RPL6是脑中最合适的组合，EF-1α和RPS18b是血液中最合适的参考基因。
High concentration of ammonia is a common issue in the aquaculture industry and often causes detrimental effects to aquatic products. Exploring expression regulation of genes involved in acute ammonia stress can help to understand the molecular mechanisms of ammonia response. Quantitative real-time PCR (qRT-PCR) with proper reference genes is an effective way to normalize the expression of target genes. To identify the most suitable reference genes forOctopus minor(Mollusca: Cephalopoda: Octopoda) under acute ammonia stress and the normal culturing, nine candidate genes were selected for the validation:OD,RPS18b,RPL29,RPS5,EF-1α,RPL6,AA4,ACT,TUB. The results showed that the stability of candidate genes varied considerably in gill, digestive gland, brain and hemolymph. Thus, the reference genes were determined separately in different tissues.RPS5andRPL6showed relatively high stability in gill, whileRPL6,TUB,RPL29andODwere four suitable reference genes in digestive gland.EF-1α,TUBandRPL6were the best combination in brain andEF-1αandRPS18bwere the most appropriate reference genes in hemolymph.