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

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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.