Overexpression of glutathione S-transferase genes in field λ-cyhalothrin resistant population of Cydia pommnella: reference gene selection and expression analysis.
Overexpression of glutathione S-transferase genes in field λ-cyhalothrin resistant population of Cydia pommnella: reference gene selection and expression analysis.
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DOI：
10.1021/acs.jafc.0c01367
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发表时间：
2020-04
影响因子：
6.1
通讯作者：
Zi-Han Wei;Miao Liu;Chao Hu;Xueqing Yang
中科院分区：
文献类型：
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作者：
Zi-Han Wei;Miao Liu;Chao Hu;Xueqing Yang
用实时荧光定量聚合酶链式反应技术分析谷胱甘肽S转移酶(GSTs)基因在耐药品系中的表达，是开展更多机制研究的关键一步，需要合适的稳定表达的参考基因。在这里，我们选择了9个常用的参考基因，并对它们的表达稳定性进行了分析。结果表明，EF-1α是所有实验集中最稳定的参考基因。EF-1α与18S、EF-1α与RPL12、EF-1α与GAPDH的组合分别足以准确地对发育阶段、组织和暴露于亚致死剂量λ-氟氯氰菊酯(LD10)的幼虫进行基因表达分析。此外，通过分析CpGSTe3、CpGSTd3和CpGSTd4在东北中国抗氯氟氰菊酯(λ)抗性群体中过表达的时空和杀虫剂诱导表达谱，验证了特定参考基因的适用性。这些基因被用来验证本研究中选择的参考基因的实用性。
Analysis of glutathione S-transferases (GSTs) gene expression in an insecticide-resistant strain of Cydia pommnella using real-time quantitative PCR is a key step towards more mechanism studies that requires suitable reference genes with stable expression. Here, nine commonly used reference genes were selected and their expression stabilities were analyzed. Results showed that EF-1α was the most stable reference gene in all of the experimental sets. The combination of EF-1α and 18S, EF-1α and RPL12, and EF-1α and GAPDH were sufficient for normalization of gene expression analysis accurately in developmental stages, tissues, and larvae exposed to sublethal dose (LD10) of λ-cyhalothrin, respectively. Additionally, the suitability of particular reference genes was verified by analyzing the spatiotemporal and insecticide-induced expression profiles of CpGSTe3, CpGSTd3, and CpGSTd4, which wereover-expressed in a λ-cyhalothrin-resistant population from northeast China. These genes were used to confer the practicability of reference genes chosen in this study.