四环素基因调节系统下小鼠VEGF基因表达效率的测定

Repression of VEGF expression in mouse tissues using tetracycline gene regulation system

为了测定四环素基因调节小鼠模型的有效性和潜在的应用价值,针对各组织中VEGF mRNA表达水平受四环素调节的效率进行了分析.从小鼠不同组织提取总RNA,进行realtime PCR分析,并利用相对定量2-△△CT方法分析这些组织中VEGF基因在mRNA表达水平上的差异,同时观察和统计小鼠组织发育情况.结果表明,转基因小鼠体内各个组织的VEFG mRNA表达水平普遍下降,尤以脂肪和卵巢最为明显.其中心脏VEFG mRNA下降53%,肝脏下降72%,肺下降50%,血小板下降49%,肾脏下降44%,脑下降55%,脂肪下降90%,子宫下降52%,卵巢下降80%.同时VEGF基因表达下降导致小鼠各个相应组织器官的发育大小出现异常.因此,四环素基因调节系统能够有效地调节转基因小鼠各个组织VEGF基因的表达水平.

Repression efficiency of mouse VEGF mRNA expression in tetracycline gene regulation system was evaluated.Total RNA was isolated from different mouse tissues including heart,liver,lung,kidney,brain,fat,uterus,ovary from double transgenic mice（VEGFteto/teto/βAtKtg/＋） with or without tetracycline treatment.VEGF messages were measured by real time quantitative PCR.The data were analyzed using the 2-△△CT method.Results show that,after removal of tetracycline,the expression of VEGF gene was down regulated by 53% in heart,72% in liver,50% in lung,49% in platelet（data published）,44% in kidney,55% in brain,90% in fat,52% in uterus,and 80% in ovary.Most organs showed significant weight loss with repression of VEGF.All above show that the tetracycline significantly regulated the VEGF mRNA expression in all the examined tissues,most dramatically down regulated in adipose tissue and ovary,accompanied with correspondingly morphological and functional disorders.