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不同生物和非生物条件下(鞘翅目:瓢虫科)实时定量聚合酶链反应分析中合适内参基因的鉴定与评价

Identification and Evaluation of Suitable Reference Genes for RT-qPCR Analysis in (Coleoptera: Coccinellidae) Under Different Biotic and Abiotic Conditions.

作者信息

Xie Jiaoxin, Liu Tinghui, Khashaveh Adel, Yi Chaoqun, Liu Xiaoxu, Zhang Yongjun

机构信息

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.

College of Plant Protections, Agricultural University of Hebei, Baoding, China.

出版信息

Front Physiol. 2021 May 17;12:669510. doi: 10.3389/fphys.2021.669510. eCollection 2021.

Abstract

Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is an accurate and convenient technique for quantifying expression levels of the target genes. Selection of the appropriate reference gene is of the vital importance for RT-qPCR analysis. is one of the most important predatory natural enemies of aphids. Recently, transcriptome and genome sequencings of facilitate the gene functional studies. However, there has been rare investigation on the detection of stably expressed reference genes in . In the current study, by using five analytical tools (Delta Ct, geNorm, NormFinder, BestKeeper, and RefFinder), eight candidate reference genes, namely, , α, , , , α, β, and , were evaluated under four experimental conditions including developmental stages, tissues, temperatures, and diets. As a result, a specific set of reference genes were recommended for each experimental condition. These findings will help to improve the accuracy and reliability of RT-qPCR data, and lay a foundation for further exploration on the gene function of .

摘要

逆转录定量聚合酶链反应(RT-qPCR)是一种用于定量目标基因表达水平的准确且便捷的技术。选择合适的内参基因对于RT-qPCR分析至关重要。[具体物种名称]是蚜虫最重要的捕食性天敌之一。最近,[具体物种名称]的转录组和基因组测序促进了基因功能研究。然而,关于[具体物种名称]中稳定表达的内参基因检测的研究很少。在本研究中,通过使用五种分析工具(Delta Ct、geNorm、NormFinder、BestKeeper和RefFinder),在包括发育阶段、组织、温度和食物在内的四种实验条件下,对八个候选内参基因,即[具体基因名称1]、α、[具体基因名称2]、[具体基因名称3]、[具体基因名称4]、α、β和[具体基因名称5]进行了评估。结果,针对每种实验条件推荐了一组特定的内参基因。这些发现将有助于提高RT-qPCR数据的准确性和可靠性,并为进一步探索[具体物种名称]的基因功能奠定基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/910a/8165390/2a6ebbbb88c9/fphys-12-669510-g001.jpg

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