通过基因表达模式识别用于半翅栉水母(鞘翅目:镍蝶科)表达分析的稳定参考基因

IF 1 Q4 GENETICS & HEREDITY
Raman Bansal, Charles S. Burks, Spencer S. Walse
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引用次数: 0

摘要

干果甲虫(Carpophilus hemipterus, Linnaeus, 1758)(鞘翅目:Nitidulidae)是果园中成熟鲜果和加工后贮藏干果的一种严重害虫。尽管半翅蓟马具有重要的经济意义且分布广泛,但却缺乏功能基因组学研究来阐明其发育生物学和分子生理学特征,以改进害虫管理。进行此类功能基因组学研究需要经过验证、稳定表达的参考基因,以便对实时定量反转录-PCR(qRT-PCR)数据进行准确归一化。在此,我们鉴定并验证了 12 个常见昆虫参考基因(RPL6、RPL4、RPL9、RPS13、RPS18、RPSA、TUB、UBE3A、RPS23、GLU、EF1A 和 RBP)在半知菌中的表达稳定性。这些基因在不同的发育阶段和性别(卵、三龄幼虫、蛹前期、蛹、雄成虫和雌成虫)以及热胁迫条件下进行了评估。候选基因的稳定性是通过软件算法 geNorm、NormFinder、BestKeeper、比较 delta-Ct 和 RefFinder 估算的。UBE3A和RPL9这两个基因在不同处理中表现出最高的稳定性,因此被提议用于归一化qRT-PCR原始数据。这项研究的结果将加强基因组学在揭示这一重要甲虫物种的基础和应用生物学方面的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Gene expression patterns to identify stable reference genes for expression analyses in Carpophilus hemipterus (L.) (Coleoptera: Nitidulidae)

Dried fruit beetle, Carpophilus hemipterus, (Linnaeus, 1758) (Coleoptera: Nitidulidae) is a serious pest of ripened fresh fruit in the orchard and dried fruit in post-processing storage. Despite the economic significance and widespread distribution of C. hemipterus, there is a lack of functional genomics research seeking to elucidate features of developmental biology and molecular physiology for improved pest management. Performing such functional genomics studies requires validated, stably expressed reference genes for accurate normalization of real-time quantitative reverse transcription-PCR (qRT-PCR) data. Here, we identified and validated 12 common insect reference genes (RPL6, RPL4, RPL9, RPS13, RPS18, RPSA, TUB, UBE3A, RPS23, GLU, EF1A, and RBP) for expression stability in C. hemipterus. These genes were assessed across developmental stages and sexes (egg, three larval instars, prepupa, pupa, adult male, and adult female) as well as thermal stress conditions. Stability of candidate genes was estimated using software algorithms geNorm, NormFinder, BestKeeper, comparative delta-Ct, and RefFinder. Two genes UBE3A and RPL9 showed the highest stability across the treatments, and thus were proposed for normalizing the qRT-PCR raw data. Results of this study will strengthen the application of genomics to reveal the fundamental and applied biological aspects in this important beetle species.

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来源期刊
Gene Reports
Gene Reports Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
3.30
自引率
7.70%
发文量
246
审稿时长
49 days
期刊介绍: Gene Reports publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses. Gene Reports strives to be a very diverse journal and topics in all fields will be considered for publication. Although not limited to the following, some general topics include: DNA Organization, Replication & Evolution -Focus on genomic DNA (chromosomal organization, comparative genomics, DNA replication, DNA repair, mobile DNA, mitochondrial DNA, chloroplast DNA). Expression & Function - Focus on functional RNAs (microRNAs, tRNAs, rRNAs, mRNA splicing, alternative polyadenylation) Regulation - Focus on processes that mediate gene-read out (epigenetics, chromatin, histone code, transcription, translation, protein degradation). Cell Signaling - Focus on mechanisms that control information flow into the nucleus to control gene expression (kinase and phosphatase pathways controlled by extra-cellular ligands, Wnt, Notch, TGFbeta/BMPs, FGFs, IGFs etc.) Profiling of gene expression and genetic variation - Focus on high throughput approaches (e.g., DeepSeq, ChIP-Seq, Affymetrix microarrays, proteomics) that define gene regulatory circuitry, molecular pathways and protein/protein networks. Genetics - Focus on development in model organisms (e.g., mouse, frog, fruit fly, worm), human genetic variation, population genetics, as well as agricultural and veterinary genetics. Molecular Pathology & Regenerative Medicine - Focus on the deregulation of molecular processes in human diseases and mechanisms supporting regeneration of tissues through pluripotent or multipotent stem cells.
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