5-脂氧化酶转基因小鼠模型的制备  

Construction of a 5-Lipoxygenase Transgenic Mice

作　　者：张梅英[1] 吴红联[1] 杨葳[1] 李兆阳[1] 董婉维[1] 周生来[1] 于洋[1] 王惟[1] 吕相川[1] 秦英[1] 郑志红[1] 王禄增[1] 

机构地区：[1]中国医科大学实验动物学部,沈阳110001

出　　处：《中国实验动物学报》2010年第1期60-64,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：辽宁省高校科研项目计划项目(2008S239);辽宁省科技厅攻关项目(2008408002-2);国家科技重大专项-药物安全评价技术平台建设项目(编号:2008ZX09305-004)

摘　　要：目的建立5-脂氧化酶(5-LO)转基因小鼠进行动脉粥样硬化的发病分子机制的研究。方法通过显微注射的方法,将5-脂氧化酶基因片段(6.8 kb)导入BDF1受精卵雄原核并移植到同期受孕的假孕母鼠输卵管中,对产出仔鼠的鼠尾组织DNA进行PCR、Southern blot检测,对9、20、24号转基因小鼠分别提取腹腔细胞、骨髓细胞及脾、肾组织总RNA和蛋白,并采用RT-PCR、Western blot方法进行转录水平检测和蛋白表达检测。结果共产生25只子代小鼠,经PCR和Southern检测获得7只阳性小鼠,经RT-PCR和Western blot检测结果表明,9、20、24号转基因小鼠腹腔细胞、骨髓细胞、脾、肾5-LO和5-脂氧化酶激活蛋白(FLAP)在RNA和蛋白水平表达均高于正常BDF1对照小鼠,且统计学分析腹腔细胞、骨髓细胞表达均具有显著差异(P<0.05)。结论成功建立5-LO转基因小鼠模型。Objective To construct a 5-lipoxygenase（5-LO） transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24 transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different（P〈0.05）.Conclusion A 5-LO transgenic mouse line has been established in this study and may be used for future study on the function of 5-LO gene.

关 键 词：5-脂氧化酶 动脉粥样硬化 转基因 

分 类 号：R711.75[医药卫生—妇产科学]