丝蛋白基因的分子设计及过表达ras1ca基因提高实用品种产丝量研究-生物化学与分子生物学专业论文.docx

丝蛋白基因的分子设计及过表达Ras1CA基因提高实用品种产丝量的研究英文摘要proteinofFib-LandGFPwas expressedsuccessfully.Inaddition,thetransgenicsilkwormhadahighsilkyieldandconvertingfoodinto silkbasedonabinaryGal4/UASsystemfromtheChineseAcademyofSciencesin Shanghai.ButitisdifficulttoapplythismethodinsericulturebecausetheRas1CAwas expressedinnon-diapausesilkworm.So,wedesignedtheGal4transgenicsilkworm hybridizedwithstockJingSongand screenedoutsomesilkworms withredfluorescenteyes,thenthenextgenerationtransgenicsilkwormwasconstantly backcrosswithJingSongtogenerateG6-Gal4diapause JingSong silkworm.wealsodesignedtheUAStransgenicsilkwormhybridizedwithstockHaoYueand screenedoutsomesilkwormswithgreenfluorescenteyes,thenthenextgeneration transgenicsilkwormwasconstantlybackcrosswithHaoYuetogenerateG6-UAS diapauseHaoYuesilkworm.ThenthetransgenicGal4 JingSong silkwormishybridized with the transgenic UAS HaoYue silkworm to get the Ras1CAspecificallyexpressed.TheresultsshowedRas1CAoverexpressionimprovedposterior silkglandweightandsilkyield11%、1.3%,confirmingsubstantialincrementofsilkproductioncapacityintransgenicsilkworm;Butfoodconsumption onlyincreased 0.57%,whichhaspotentialinsericulture.As measured by quantitative real-time PCR,Ras1CAmRNAlevelintheposteriorsilkglandofFil-Gal4/UAS-Ras1CAsilkwormwas4.08foldhigherthanincontrols.TheseresultsensuredRas1CA issuccessfullyoverexpressedintheposteriorsilkgland.Keywords:silkworm;genetargeting;Gfp;fibroinlightchaingene;fusion expression; Ras1CA;Gal4/UAStransgenicsystem;posteriorsilkgland;silkyield.Writtenby:LiJingzhiSupervisedby:GONGChengliangIII目录第一部分文献综述1第一章基因打靶技术的研究进展11基因打靶技术的发展12 基因打靶家蚕研究的现状4参考文献5第二章抗菌肽的研究进展91抗菌肽的抗菌机制92抗菌肽的生物活性103抗菌肽应用前景104 家蚕抗菌肽基因研究进展12参考文献14第三章Gal4/UAS双元系统在转基因技术中研究进展161PiggyBac介导的Gal4/UAS双转基因系统182Gal4转基因系和UAS靶基因系的构建193Gal4/UAS的应用20参考文献21第二部分研究报告24第四章研究目的与内容241研究目的及意义242研究策略与内容253 技术路线25第五章实验材料与方法271 实验材料272 常用仪器303 实验方法30参考文献36第六章Fib-L融合GFP及抗菌肽的转基因家蚕的设计与制作361材料与方法392结果与分析423讨论47参考文献48第七章基于GAL4/UAS转基因系统提高现行家蚕品种产丝量的研究511 材料与方法522 结果与分析533 讨论59参