5-脂氧化酶转基因小鼠模型的制备
5-지양화매전기인소서모형적제비
Construction of a 5-Lipoxygenase Transgenic Mice
  • 万方数据
    中国实验动物学报 중국실험동물학보
    ACTA LABORATORIUM ANIMALIS SCIENTIA SINICA
    2010年 1期 60-64 ,共5页

    张梅英%吴红联%杨葳%李兆阳%董婉维%周生来%于洋%王惟%吕相川%秦英%郑志红%王禄增

    장매영%오홍련%양위%리조양%동완유%주생래%우양%왕유%려상천%진영%정지홍%왕록증

    5-脂氧化酶%动脉粥样硬化%转基因 5-脂氧化酶%動脈粥樣硬化%轉基因
    5-지양화매%동맥죽양경화%전기인
    5-lipoxygenase%Atheroselerosis%Transgenic mice
    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.
    목적 건립5-지양화매(5-LO)전기인소서진행동맥죽양경화적발병분자궤제적연구.방법 통과현미주사적방법,장5-지양화매기인편단(6.8 kb)도입BDF1수정란웅원핵병이식도동기수잉적가잉모서수란관중,대산출자서적서미조직DNA진행PCR、Southern blot검측,대9、20、24호전기인소서분별제취복강세포、골수세포급비、신조직총RNA화단백,병채용RT-PCR、Western blot방법진행전록수평검측화단백표체검측.결과 공산생25지자대소서,경PCR화Southern검측획득7지양성소서,경RT-PCR화Western blot검측결과표명,9、20、24호전기인소서복강세포、골수세포、비、신5-LO화5-지양화매격활단백(FLAP)재RNA화단백수평표체균고우정상BDF1대조소서,차통계학분석복강세포、골수세포표체균구유현저차이(P<0.05).결론 성공건립5-LO전기인소서모형.
    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.9,20,24transgenic 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 transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.
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