The identification of Cytochrome P450s involved in the biosynthesis of ecdysteroid hormon and their gene expression systems

参与蜕皮激素生物合成的细胞色素P450及其基因表达系统的鉴定

• 批准号: 14560046
• 负责人: TOMITA Takashi
• 金额: $ 2.3万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560046/
• 关键词: Ecdysteroid; CYP302A1; CYP315A1; CYP312A1; CYP6A19; Cytochrome P450; Drosophila melanogaster; 脱皮ホルモン; ゲノム; CYP302Al; CYP315Al; CYP312Al

Using cDNA array techniques, the transcriptional levels of Drosophila cytochrome P450 (P450) genes were compared between male and female flies. Of the 86 P450s,15 isoforms were picked up and the levels of transcription were confirmed by the real time quantitative RT-PCR. Cyp315a1 and Cyp302a1,which had been reported as P450s involved in the ecdysteroid biosynthesis, were included in the 15 isoforms and expression of these P450s was 8.6-and 7.9-fold higher in females than in males, respectively. In addition, we confirmed that expression of Cyp312a1 was 82-fold higher in adult males than females. This gene expression was observed mostly in the abdomen and its transcription level gradually increased from pupal stage and peaked in the 5-day-old adult. Furthermore, the male specific expression of Cyp312a1 was universally observed in three Drosophila strains originated from the USA (Oregon R), China (Canton S), and Japan (HKJ), suggesting possible involvement of this P450 in significant endogenous catalytic reaction(s). This is the first report of a P450 enzyme being predominantly expressed in male Drosophila.

利用cDNA阵列技术，比较了果蝇细胞色素P450（P450）基因在雄性和雌性果蝇之间的转录水平。在86个P450基因中，筛选出15个亚型，并通过真实的实时定量RT-PCR证实其转录水平。Cyp 315 a1和Cyp 302 a1被报道为参与蜕皮激素生物合成的P450，它们被包括在15种亚型中，这些P450在雌性中的表达分别是雄性的8.6倍和7.9倍。此外，我们证实Cyp 312 a1的表达在成年男性中比女性高82倍。该基因主要在腹部表达，其转录水平从蛹期开始逐渐增加，在5日龄成虫时达到高峰。此外，Cyp 312 a1的雄性特异性表达普遍存在于来自美国（俄勒冈州R）、中国（Canton S）和日本（HKJ）的三个果蝇品系中，表明该P450可能参与重要的内源性催化反应。这是第一个报告的P450酶主要在雄性果蝇中表达。

项目成果

S.Kasai, T.Tomita: "Male specific expression of a cytochrome P450 (Cyp312al) in Drosophila melanogaster"Biochemical and Biophysical Research Communications. 300(4). 894-900 (2003)

S.Kasai、T.Tomita：“果蝇中细胞色素 P450 (Cyp312al) 的雄性特异性表达”生物化学和生物物理研究通讯。

Kasai S, Tomita T.: "The analysis of P450 gene expression with cDNA array : In search for novel insecticidal targets and the study of insecticide-resistance mechanisms."Journal of Pesticide Science (in Japanese). 28(4). 473-478 (2003)

Kasai S、Tomita T.：“用 cDNA 阵列分析 P450 基因表达：寻找新的杀虫靶点和杀虫剂抗性机制的研究。”农药科学杂志（日文）。

Kasai S.: "The study on cytochrome P450s as pyrethroid-resistance factors"Journal of Pesticide Science (in Japanese). (in press). (2003)

Kasai S.：“细胞色素P450作为拟除虫菊酯抗性因子的研究”农药科学杂志（日文）。

Kasai S, Tomita T.: "Male specific expression of a cytochrome P450 (Cyp312a1) in Drosophila melanogaster."Biochemical and Biophysical Research Communications. 300(4). 894-900 (2003)

Kasai S、Tomita T.：“细胞色素 P450 (Cyp312a1) 在果蝇中的雄性特异性表达。”生物化学和生物物理研究通讯。

葛西真治, 冨田隆史: "cDNAアレイ法によるチトクロムP450発現の解析:殺虫剤新規作用点の探索と抵抗性機構の解明に向けて"日本農薬学会誌. 28(4). 473-478 (2003)

Shinji Kasai、Takashi Tomita：“使用 cDNA 阵列方法分析细胞色素 P450 表达：寻找杀虫剂的新作用位点并阐明抗性机制”日本农药学会杂志 28(4) (2003)。