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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lox P位点的转基因小鼠。在FKBP38基因位置携带lox P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:(1)FKBP38肝脏特异敲除小鼠FKBP38~(-/-)肝脏中FKBP38基因的m RNA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)。(2)FKBP38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)。(3)FKBP38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。(4)FKBP38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m RNA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。
Objective: To construct liver specific knockout mice with FKBP38 (FK506 Binding Protein 38) gene. Methods: The transgenic mice carrying lox P site on FKBP38 were constructed by embryo injection. On the basis of the loxP site in the FKBP38 gene locus, Alb-Cre, which is specifically expressed in hepatic parenchymal cells, mediates FKBP38 conditional knockout to obtain a liver-specific knockout mouse model of FKBP38 gene Alb-Cre: FKBP38 ~ (fl / fl). At the same time, FKBP38 specific knockout mice were identified. Results: (1) The mRNA level of FKBP38 gene in FKBP38 ~ (-) - / - liver of FKBP38 liver-specific knockout mice was statistically different from that of wild-type mice of the same age (P <0.001). (2) The protein expression of FKBP38 in FKBP38 liver-specific knockout mice FKBP38 ~ (- / -) liver was statistically different from that in wild-type mice of the same age (P <0.001). (3) There was no significant difference in the level of transcription and translation of FKBP38 liver-specific knockout mouse FKBP38 ~ (- / -) liver, phosphorylation level of p70 S6K slightly increased and phosphorylation level of 4EBP-1 slightly decreased . (4) There was no differentiated expression of Bcl-2 in FKBP38 liver-specific knockout mice FKBP38 ~ (- / -) liver. CONCLUSION: FKBP38 liver-specific knockout mouse FKBP38 ~ (- / -) liver mRNA and protein of FKBP38 gene is basically not expressed, suggesting successful construction of liver specific knockout mice FKBP38 gene.