Identification and cloning of JH receptor

JH受体的鉴定与克隆

• 批准号: 10460020
• 负责人: CHINZEI Yasuo
• 金额: $ 8.38万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10460020/
• 关键词: Juvenile hormone; metamorphosis; transcription regulation; cDNA library; cyanoprotein; リセプター; エクダイソン; 転写因子; 変態関連遺伝子; ルシフェラーゼ; EcR; USP; 遺伝子発現; 転写調節因子; シアノプロテイン

Juvenile hormone (JH) regulates a wide range of insect physiological phenomena such as metamorphosis, diapause and polymorphysm, which are very important for insect development and complete of life cycle. However JH mode of action is still unknown, particularly JH receptor and signal transduction in the target cells are not elucidated yet.In this study we aimed to make clear a moleculara level of JH action including JH receptor and the regulation of gene expression through the cloning of JH responsive gene and their responsive transcription factor, and identification and cloning of JH receptor.We studied the cloning of JH responsive transcription factor (JHRTF), by the experiments combined that showed below, using JH responsive element (JHRE) of cyanoprotein (CP) gene and JH treated fat body cDNA library, in the bean bug Riptortus clavatus.(1) JHRTF is screened by south-Western method and phage-display method from cDNA library transformed In the expression vector or Phase by using labelled JHRE as a probe.(2) JHRTF Is screened by one-hybrid method, Introducing DNA constract which constructed with JHR E of CP, and cDNA-Gal4 fusion library Into yeast cells.(3) JHRTF Is screened by two-hybrid method, using cDNA-jGla4 fusion library and Gla4 fusion DNA construct with of known transcription factor.We have tried to study the above experiments, It was unsuccessful, and we could not get the JHRDT clone as a result so far.

保幼激素(JH)调节昆虫的变态、滞育、多态等多种生理现象，对昆虫的发育和生活史的完成具有重要意义。然而，JH的作用机制尚不清楚，尤其是JH受体及其在靶细胞中的信号转导尚不清楚。本研究旨在通过克隆JH反应基因及其反应转录因子，以及JH受体的鉴定和克隆，明确包括JH受体在内的JH作用的分子水平及其对基因表达的调控。本研究结合以下实验，利用氰蛋白(CP)基因的JH反应元件(JHRE)和JH处理后的脂肪体cDNA文库，对JH反应转录因子(JHRTF)的克隆进行了研究。(1)以标记的JHRE为探针，用西南法和噬菌体展示法从表达载体或相中转化的cDNA文库中筛选出JHRTF。(2)用单杂交方法筛选JHRTF，将与CP的JHR E构建的DNA结构与cDNAGal4融合文库导入酵母细胞中。(3)用双杂交方法筛选JHRTF，利用已知转录因子的Gla4融合文库和Gla4融合文库进行筛选，我们尝试了上述实验，但均未成功，未能获得JHRDT克隆。

项目成果

Hubert Woitasek et al.: "Analysis of the involvement of the 3'-untraslated regions in regulating mRNA stability for vitellogenin, cyanoprotein a and"Insect Biochem. Mol. Biol.. (in press). (2002)

Hubert Woitasek 等人：“分析 3-非翻译区域参与调节卵黄原蛋白、氰基蛋白 a 和 Insect Biochem 的 mRNA 稳定性。”

T.Shinoda, et al.: "Characterization of a recombinant insect chitinase and a baculovirus chitinase."Eur.J.Biochem.. (in press).

T.Shinoda 等人：“重组昆虫几丁质酶和杆状病毒几丁质酶的表征。”Eur.J.Biochem..（正在印刷中）。

M.Hirai, et al.: "A juvenile hormone-repressibletransferrin-like protein from the bean bug, Riptortus clavatus : cDNA sequence analysis ans protein identification during diapause and vitellogenesis."Arch.Insect Biochem.Physiol.. 44. 17-26 (2000)

M.Hirai 等人：“来自豆虫 Riptortus clavatus 的保幼激素抑制转铁蛋白样蛋白：滞育和卵黄发生期间的 cDNA 序列分析和蛋白鉴定。”Arch.Insect Biochem.Physiol.. 44. 17-26

Y.Furuta, et al.: "Peroral infectivity of non-occluded viruses of Bombyx mori nucleopolyhedrovirus and polyhedrin-negative recombinant"J. Gen. Virol.. 82. 307-312 (2001)

Y.Furuta 等人：“家蚕核多角体病毒和多角体蛋白阴性重组体的非包涵体病毒的经口感染性”J.

Toru Kariu, et.al.: "Essential function of MAEBL in malarial sporozoite infection of the mosquito salivary gland"J.Exp.Med.. (in press). (2002)

Toru Kariu 等人：“MAEBL 在蚊子唾液腺疟疾子孢子感染中的基本功能”J.Exp.Med..（出版中）。