脑部特异表达rtTA调控元件转基因小鼠的构建  

Construction of forebrain-specific rtTA expression transgenic mouse

作　　者：章蓉[1] 孙强[1] 徐桂芳[1] 董娟[1] 金宇娟[1] 江楠[1] 

机构地区：[1]华东师范大学脑功能基因组学研究所,上海200062

出　　处：《扬州大学学报（农业与生命科学版）》2008年第1期16-19,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家"十五"科技攻关项目(2001BA70113)

摘　　要：为获得一种可调控基因表达系统,避免传统转基因在胚胎发育早期表达的毒性作用或致死作用以及基因过度表达而造成难于分析的复杂多表型,应用分子生物学方法将四环素反式激活子rtTA克隆到仅在脑部能被激活的钙调蛋白依赖性蛋白激酶CaM kinaseⅡα亚基启动子之后,将其通过显微注射方法注入FVB小鼠受精卵雄原核中获得四环素调控系统中反应组分的转基因工具鼠。经PCR及Southern检测,最终得到3只阳性小鼠(1公2母),表达检测结果显示,其中仅2只(1公1母)后代检测到rtTA表达产物。结果表明已成功构建四环素调控系统中的转基因工具鼠,为今后研究其他脑部基因功能提供便利。Conditional gene expression may help to avoid not only blocking analysis of phenotype caused by toxic or lethal effects in embryonic development early time, but also complicating phenotype caused by ubiquitous overexpression, so we utilized the benefits of an optimized tet-on system to construct p279-rtTA with molecular biology method, then it was transferred into FVB mice by pronucleus microinjection to produce transgenic mouse, to this constructed forebrain-specific rtTA transgenic tool mouse. Through PCR and Soutnern blot analysis, three independent lines of transgenic mice carrying the CaM kinase IIalpha promoter-rtTA gene was generated （1 ♂ 2 ♀ ） from 179 mice. The expression of rtTA was detected in the brain of F1 carrying CaM kinase Ⅱ alpha promoter-rtTA gene, and not in the other tissues. The results indicated that the construction of the CaM kinase Ⅱ alpha promoter-rtTA transgenic tool mice was successful. It will be useful tool for future genetic studies of brain functions and researches of gene treatment.

关 键 词：小鼠 转基因 RTTA CaM kinaseⅡ 

分 类 号：Q786[生物学—分子生物学] S858.91[农业科学—临床兽医学]