| 王 帅,赖姨梅,林 艳,李晓曦,赵子建.FKBP38肝脏特异敲除小鼠模型的构建[J].,2017,17(26):5001-5006 |
| FKBP38肝脏特异敲除小鼠模型的构建 |
| Construction of a Mouse Model of Hepatocyte Cell-specific Disruption of the FKBP38 Gene |
| 投稿时间：2017-03-30 修订日期：2017-04-20 |
| DOI：10.13241/j.cnki.pmb.2017.26.001 |
| 中文关键词: FKBP38 肿瘤 条件性敲除 Cre/loxP重组酶系统 |
| 英文关键词: FKBP38 Tumor Conditional knock out Cre/loxP recombinase system |
| 基金项目:国家重点基础研究发展规划(973计划)（2013CB945202）；国家自然科学基金项目（81372798）；江苏省自然科学基金项目（BK20130059） |
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| 中文摘要: |
| 摘要 目的：构建FKBP38(FK506 Binding Protein 38)基因肝脏特异敲除小鼠。方法：利用胚胎注射法构建在FKBP38上携带loxP位点的转基因小鼠。在FKBP38基因位置携带loxP位点的小鼠的基础上，以肝脏实质细胞特异性表达的Alb-Cre介导FKBP38条件性敲除，以获得FKBP38基因肝脏特异敲除小鼠模型Alb-Cre:FKBP38 fl/fl 。同时对FKBP38特异性敲除鼠进行鉴定。结果：①FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中FKBP38 基因的mRNA水平相对于同年龄同窝野生型小鼠具有统计学差异（P＜0.001）。②FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中FKBP38 基因的蛋白表达水平相对于同年龄同窝野生型小鼠具有统计学差异（P＜0.001）。③FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中，转录和翻译相关蛋白水平未见显著差异，p70 S6K的磷酸化水平轻微上调，4EBP-1的磷酸化水平有轻微下调。④FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中，凋亡相关蛋白Bcl-2未见差异化表达。结论：FKBP38肝脏特异敲除小鼠FKBP38-/-肝脏中，FKBP38 基因的mRNA和蛋白基本不表达，提示成功构建FKBP38基因肝脏特异敲除小鼠。 |
| 英文摘要: |
| ABSTRACT Objective: To build the model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver. Methods: Transgenic mouse whose FKBP38 gene was flanked with loxP was constructed by embryo microinjection. The FKBP38 gene was deleted by breeding mice harboring two loxP sites in FKBP38 (FKBP38 fl/fl ) with the mice bearing the expression of Cre recombinase mice driven by an album promoter. Afterward, the genotype of FKBP38 conditional knockout mice was analyzed. Results: ①Relative hepatic FKBP38 mRNA levels showed significant difference between FKBP38 conditional knockout mice (FKBP38-/-) and wild type(P＜0.001).②Relative hepatic FKBP38 protein expression levels of FKBP38 conditional knockout mice (FKBP38-/-) were significantly different with wild type(P＜0.001).③Relative phosphorylation of hepatic p70 S6K and 4E-BP-1 protein of FKBP38 conditional knockout mice (FKBP38-/-) showed no significant difference, with slight decrease in phosphorylation of 4E-BP-1, compared with wild type. ④No significant difference in expression of hepatic Bcl-2 between FKBP38-/- and wild type. Conclusion: The mouse model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver is successfully built. |
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