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蜚蠊寄生蜂(拉策堡)定量实时PCR分析中内参基因的筛选与验证

Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis in Cockroach Parasitoid (Ratzeburg).

作者信息

Dong Renke, Cao Fengming, Yu Jincong, Yuan Yuan, Wang Jiahui, Li Zining, Zhu Chunxue, Li Sheng, Li Na

机构信息

Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China.

出版信息

Insects. 2024 Sep 3;15(9):668. doi: 10.3390/insects15090668.

Abstract

Parasitoid wasps play a crucial role in the efficient control of pests, a substantial menace to human health and well-being. (Ratzeburg) stands out as the most effective egg parasitoid wasp for controlling American cockroaches, but accurate and stable reference genes for quantitative real-time polymerase chain reaction of genes are still lacking. In this study, we assessed seven candidate nuclear genes, including (), (), (), (), (), (), and () of . By analyzing expression stability with four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), as well as comprehensive ranking with RefFinder, we identified as the most stable reference gene for the larval, pupal, female adult, and male adult stages. Subsequently, we estimated the transcript levels of vitellogenin () and cuticle protein () after normalization with across various developmental stages. Significantly higher expression levels of and were observed in pupae and female adults, respectively, consistent with previous findings in other insects. This study offers a reliable reference gene for normalizing transcription levels of genes.

摘要

寄生蜂在有效控制害虫方面发挥着关键作用,害虫对人类健康和福祉构成重大威胁。(拉策堡)黄蜂是控制美洲大蠊最有效的卵寄生蜂,但用于基因定量实时聚合酶链反应的准确且稳定的参考基因仍然缺乏。在本研究中,我们评估了七个候选核基因,包括(基因名称1)、(基因名称2)、(基因名称3)、(基因名称4)、(基因名称5)、(基因名称6)和(基因名称7)。通过使用四种算法(Delta Ct、geNorm、NormFinder和BestKeeper)分析表达稳定性,并通过RefFinder进行综合排名,我们确定(基因名称)是幼虫、蛹、成年雌性和成年雄性阶段最稳定的参考基因。随后,我们在用(参考基因名称)标准化后,估计了不同发育阶段卵黄原蛋白(基因名称)和表皮蛋白(基因名称)的转录水平。分别在蛹和成年雌性中观察到(基因名称)和(基因名称)明显更高的表达水平,这与之前在其他昆虫中的发现一致。本研究为标准化(目标基因名称)基因的转录水平提供了一个可靠的参考基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b28f/11431952/99e77d2d33f2/insects-15-00668-g001.jpg

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