【作者】 王春生； 宁方勇； 李昊； 梁洋； 朴善花； 安铁洙；

【Author】 WANG Chun-sheng,YANG Yue-fei,LI Hao,QU Jun-jie,LUO Fang,AN Tie-zhu WANG Chun-sheng,AN Tiezhu College of Life Science,Northeast Forestry University,harbin,150040,China

【机构】 东北林业大学生命科学学院；

【摘要】 为了筛选在小鼠毛囊中具有高表达活性的内源启动子,本研究以小鼠基因组为模板,克隆得到超高硫角蛋白基因启动子UHS,将其分别与pβgal-Basic和pAcGFP1-N1载体连接,构建了重组真核表达载体。采用阳离子脂质体法转染胎鼠组织块,分析其表达活性。结果显示,转染后48h,在蓝光激发条件下可以检测到GFP在小鼠毛囊区高表达,转染96h后,表达减弱:此外,转染后48h,βgal染色结果显示在皮肤快的毛囊区存在蓝色点状区域,表明UHS启动子在小鼠毛囊中具有表达特异性。更多还原

【Abstract】 The Keratin is main structure albumen of the mammal wool,which is the most important part of skin,hair and nail,etc.In order to detect the activity of Ultra high sulfur Keratin promoter in mouse skin,our experiment regards the genomic DNA from the mouse as the template,through molecular biologic methods to clone the mouse Ultra high sulfur Keratin promoter UHS,identifying after joint pMD18 simple vector,then insert it into pAcGFP-N1 plasmid which had been excised the CMV promoter,after PCR identifying,then recombined a eukaryotic expression vector.At the same tine,the promoter was linked to pβgal-Basic,and recombination plasmid U-GFP and U-β-gal were obtained respectively.The mouse skin tissue piece was transfected by cationic liposome method with this plasmids.The results showed that:after transfected 48h,GFP was detected at mouse hair follicle region under blue blaze light,and the expression weaken after 96h.Moreover, after transfected 48h,βgal strain result showed there were blue dots at mouse hair follicle region,and indicted the promoter was tissue-specific promoter at mouse hair follicle.更多还原