胰岛细胞特异性敲除基因appl1小鼠模型的制备.doc

尊敬的《上海医学》编辑老师： 您好！首先非常感谢您及评审专家对我文章的审修，已按照修改意见进行了修改，以下是对修改意见的具体答复，希望 1.“胰岛细胞特异性敲除APPL1基因”和“β细胞APPL1特异性敲除APPL1基因”这两种表达方法应该统一；在文章中已做修改 2.从图3可以看出，β细胞APPL1敲除小鼠脂肪组织APPL1的表达显著高于野生型，原因何在?我们APPL1flox/flox和β-APPL1KO两组小鼠多例样本并未发现3.该研究目的是建立β细胞APPL1基因敲除小鼠模型，因此在表型分析方面应多关注β细胞功能，至少要提供血胰岛素水平方面的数据，而不是仅仅提供血糖数据。我们研究组已经研究了APPL1flox/flox小鼠的空腹血胰岛素水平差异 4. 如可以请引用《上海医学》杂志和《中国实用内科杂志》的参考文献各一条 在文章中我已做 此致 敬礼 李晓雯 2015.1．30 胰岛细胞特异性敲除基因APPL1小鼠模型的制1 李羚1 林紫薇1 孙赟1 陈辰2 王琛1* 贾伟平1 【摘要】 目的 转接蛋白APPL1方法 采用囊胚显微注射法制备嵌合小鼠APPL1胰岛特异性敲除小鼠。Western blot检测蛋白表达结果 胰岛细胞特异性敲除基因小鼠模型APPL1flox/flox小鼠相比，胰岛细胞特异性敲除基因 blot从蛋白水平印证敲除小鼠胰岛中的APPL1蛋白无表达。 结论 胰岛细胞特异性敲除基因小鼠模型制。 【关键词】APPL1；Establishment of APPL1 conditional knock out model in mice islet LI Xiaowen1, LI Ling1, LIN Zeiwei1, SUN Yun1, CHEN Chen2 , WANG Chen1*, JIA Weiping1. 1Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai 200233, China 2 Shanghai Research Center for Model Organisms, Shanghai 201210, China Corresponding author: WANG Chen, wangchen@sjtu.edu.cn 【Abstract】 Objective To discuss the method of model in islet. Methods Mouse embryonic stem (ES) cells were targeted knockout of APPL1 by the homologous recombination vector, and screened .The APPL1-knockout embryonic stem cells were microinjected into blastula of C57BL/6J mice.F1 hybrid mice were bred to obtain mouse aggregation chimeras. The conditional KO mice were generated by cross-breeding APPL1 floxed mice with mice expressing Cre in islets. Western blot was conducted to measure protein expression. Results Mice with conditional?APPL1?knockout (KO) in islets were generated. Compared with APPL1flox/flox mice, APPL1?conditional knockout? in islets does not affect the mice weight, fasting plasma glucose and fasting insulin levels, no APPL1 protein expression was found in APPL1 KO mice islets with western blot. Conclusions The conditional APPL1-knockout mouse model is successfully