The cloning of hormone sensitive gene controling insect function by using gene technolog

利用基因技术克隆控制昆虫功能的激素敏感基因

• 批准号: 07660068
• 负责人: KAWASAKI Hideki
• 金额: $ 1.47万
• 依托单位: Utsunomiya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660068/
• 关键词: Bombyx mori; wing disc; beta-tubulin; ecdysteroid; ditterential bybridization; metamorphosis; insect function; ディファレンシャルスクリーニング

The cDNA library of wing disc and fat body were constructed. Those of wing discs were made from mRNAs induced or not induced by ecdysteroid. These cDNA Libraries were ligated to lambdaZap vector system. Bombyx genomic library were constructed and ligated to lambdaDASH system. cDNA probes were made from different stage and hormone induced or not induced mRNAa of wing discs and differential screening was operated. Putative stage specific and ecdysone-inducible 5 cDNA clones were obtained. These clones will be examined for northern hibridization whether these are stage specific or ecdysone-inducible. Some peptides were induced stage specifically or by addition of hormone and were detected by using 2-dimensional gel electrophoresis autoradiography and the results are in preparation of publication.Two primers were constructed among very conservative DNA region of beta-tubulin gene and PCR amplification was done. Putative tubulin cDNA were amplified. cDNA libraries of wing discs were screened by using this PCR products as non-RI probe. Two types of beta-tubulin genes were cloned. One of them were induced stage specifically and by ecdysone, now they are in analysis of gene organization.

构建了翼盘和脂肪体的cDNA文库。翼盘是由蜕皮类固醇诱导或非诱导的 mRNA 制成的。将这些 cDNA 文库连接至 lambdaZap 载体系统。构建了家蚕基因组文库并将其连接至 lambdaDASH 系统。从不同阶段和激素诱导或非诱导的翼盘mRNAa制备cDNA探针并进行差异筛选。获得了假定的阶段特异性和蜕皮激素诱导的5个cDNA克隆。将检查这些克隆的北方杂交，无论它们是阶段特异性的还是蜕皮激素诱导的。部分肽段经过特异性诱导或添加激素诱导，采用二维凝胶电泳放射自显影技术进行检测，结果正在准备发表。在β-微管蛋白基因非常保守的DNA区域中构建两条引物，进行PCR扩增。扩增推定的微管蛋白cDNA。以此PCR产物作为non-RI探针筛选翼盘cDNA文库。克隆了两种类型的β-微管蛋白基因。其中之一是由蜕皮激素特异性诱导的，现在正在进行基因组织分析。

项目成果

川崎秀樹: "カイコ突然変異無翅における5齢期の翅の形態形成" 日本蚕糸学雑誌. 66. 48-53 (1997)

川崎秀树：“突变无翅蚕第五龄翅膀的形态发生”，日本蚕丝学杂志 66. 48-53 (1997)。

HIDEKI KAWASAKI: "Wing disc horphosenesis of flmutant (flugellose) of Bombyx mori during the fifth larval instar." J.Sericultural Science of Japan. 66(1). 48-53 (1997)

HIDEKI KAWASAKI：“家蚕第五龄幼虫期间翼盘突变体（flugellose）的翅盘分裂。”

Hideki Kawasaki: "Transition from larva to pupa : morphogenesis, cell proliteration and gene expression in Bombyx wing disc." Invertebrate Peproduction & Pevelopment. (in press).

Hideki Kawasaki：“从幼虫到蛹的转变：家蚕翅盘的形态发生、细胞增殖和基因表达。”

Hideki Kawasaki: "Transition from larva to pupa : morphogenesis,cell proliteration and gene expression in Bombyx wing disc." Invertebrate Reproduction & Development. (in press).

Hideki Kawasaki：“从幼虫到蛹的转变：家蚕翼盘的形态发生、细胞增殖和基因表达。”