转β-甘露聚糖酶基因manA小鼠的制备  

Generation of beta-mannanase manA transgenic mice

作　　者：李紫聪[1] 张茂[2] 许惠[1] 刘德武[1] 吴珍芳[1] 

机构地区：[1]华南农业大学动物科学学院,广东广州510642 [2]龙岩学院生命科学学院,福建龙岩364012

出　　处：《广东农业科学》2013年第14期144-146,150,共4页Guangdong Agricultural Sciences

基　　金：广东省科技计划项目(2011A020901001;2011A020102003);国家自然科学基金青年项目(31101689)

摘　　要：为了获得β-甘露聚糖酶基因manA的转基因小鼠,从黑曲霉中克隆得到β-甘露聚糖酶基因manA,进行体外表达检测甘露聚糖酶活性后,将此基因插入到含有猪腮腺分泌蛋白(PSP)基因启动子的表达载体pPSPBGPneo中,得到在腮腺组织特异表达β-甘露聚糖酶基因manA的载体pPSP-manA,总长为16.3 kb,将其进行线性化后回收得到高质量DNA片段,通过显微注射得到17只原代小鼠,进行PCR和Southern blot检测发现有6只阳性转基因小鼠,表明转β-甘露聚糖酶基因manA小鼠制备成功。In order to obtain the transgenic mice of beta-mannanase, mariA gene of beta-mannanase from Aspergillus niger was cloned and the activity of beta-mannanase expressed was detected. Then the manA gene was inserted into the expression vector pPSPBGPneo that contain the promoter of pig parotid secretory protein （PSP）gene to obtain the new vector of pPSP-manA, which had special expression in pig parotid. 16.3 kb long of the new vector was then linearized and microinjected into the mice embryo to gain 17 primary mice. Finally, 6 positive transgenic mice that contain the mcmA gene of beta-mannanase were obtained successfully after PCR and Southern blot detection.

关 键 词：Β-甘露聚糖酶 manA基因 转基因小鼠 

分 类 号：S813.3[农业科学—畜牧学]