Alcohol dehydrogenase 5 (ADH5) is a member of medium‐chain dehydrogenase/reductase family and takes part in cellular formaldehyde and S‐nitrosoglutathione metabolic network. 2‐tridecanone (2‐TD) is a toxic compound in many Solanaceae crops to defend against a variety of herbivory insects. In the broader context of insect development and pest control strategies, this study investigates how a new ADH5 from Helicoverpa armigera (HaADH5) regulates the expression of CYP6B6, a gene involved in molting and metamorphosis, in response to 2‐TD treatment. Cloning of the HaADH5 complementary DNA sequence revealed that its 1002 bp open reading frame encodes 334 amino acids with a predicted molecular weight of 36.5 kD. HaADH5 protein was purified in the Escherichia coli Transetta (pET32a‐HaADH5) strain using a prokaryotic expression system. The ability of HaADH5 protein to interact with the 2‐TD responsive region within the promoter of CYP6B6 was confirmed by an in vitro electrophoretic mobility shift assay and transcription activity validation in yeast. Finally, the expression levels of both HaADH5 and CYP6B6 were found to be significantly decreased in the midgut of 6th instar larvae after 48 h of treatment with 10 mg/g 2‐TD artificial diet. These results indicate that upon 2‐TD treatment of cotton bollworm, HaADH5 regulates the expression of CYP6B6 by interacting with its promoter. As HaADH5 regulation of CYP6B6 expression may contribute to the larval xenobiotic detoxification, molting and metamorphosis, HaADH5 is a candidate target for controlling the growth and development of cotton bollworm.

中文翻译：

---

棉铃虫乙醇脱氢酶 5 与 CYP6B6 启动子相互作用以响应 2-十三酮

酒精脱氢酶 5 (ADH5) 是中链脱氢酶/还原酶家族的成员，参与细胞甲醛和 S-亚硝基谷胱甘肽代谢网络。2-十三烷酮 (2-TD) 是许多茄科作物中的一种有毒化合物，可抵御多种食草昆虫。在昆虫发育和害虫防治策略的更广泛背景下，本研究调查了棉铃虫 (HaADH5) 的新 ADH5 如何调节 CYP6B6 的表达，CYP6B6 是一种参与蜕皮和变态的基因，以响应 2-TD 处理。HaADH5 互补 DNA 序列的克隆显示其 1002 bp 开放阅读框编码 334 个氨基酸，预测分子量为 36.5 kD。使用原核表达系统在大肠杆菌 Transetta (pET32a-HaADH5) 菌株中纯化 HaADH5 蛋白。HaADH5 蛋白与 CYP6B6 启动子内的 2-TD 反应区相互作用的能力通过体外电泳迁移率变化测定和酵母转录活性验证得到证实。最后，发现在用 10 mg/g 2-TD 人工饲料处理 48 小时后，HaADH5 和 CYP6B6 在 6 龄幼虫中肠中的表达水平显着降低。这些结果表明，对棉铃虫进行 2-TD 处理后，HaADH5 通过与其启动子相互作用来调节 CYP6B6 的表达。由于 HaADH5 对 CYP6B6 表达的调节可能有助于幼虫外源性解毒、蜕皮和变态，因此 HaADH5 是控制棉铃虫生长发育的候选靶点。