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目的:Lin28是一种高度保守的RNA结合蛋白,对生物体的生命活动具有重要的调节作用。为了研究人Lin28A与Lin28B基因的功能,我们构建了分别过表达人Lin28A或Lin28B基因的两种转基因小鼠,表型分析发现小鼠体重变化,为研究Lin28A和Lin28B基因的生物学功能提供模型动物。方法:分别将人类Lin28A与Lin28B cDNA插入PCAG启动子下游,构建Lin28A与Lin28B转基因表达载体,通过显微注射方法,分别建立Lin28A转基因小鼠与Lin28B转基因小鼠。PCR鉴定转基因小鼠的基因型,筛选出人类Lin28A与Lin28B表达的小鼠,统计两种转基因小鼠体重变化。结果:①经PCR鉴定与公司测序证明Lin28A和Lin28B两种载体构建成功。②PCR鉴定小鼠基因型,筛选出可以特异性表达人Lin28A或Lin28B的转基因小鼠,并比较转基因小鼠与同窝野生型小鼠体重,发现两种转基因小鼠体重均大于同窝野生型小鼠,说明在小鼠体内过表达Lin28A或Lin28B均能引起小鼠体重增加。结论:人Lin28A与Lin28B在转基因小鼠体内能正常表达,并且能发挥生物学功能。又因为Lin28A和Lin28B在小鼠体内过表达会引起小鼠体重增加,我们推测其可能通过影响小鼠糖代谢过程引起小鼠体重改变。构建的Lin28A和Lin28B的转基因小鼠为进一步研究Lin28A和Lin28B在人新陈代谢、生长和发育过程中的作用提供了动物模型。
Objective: Lin28 is a highly conserved RNA-binding protein that plays an important regulatory role in the life activities of living organisms. To investigate the function of human Lin28A and Lin28B genes, we constructed two transgenic mice overexpressing human Lin28A or Lin28B respectively. Phenotypic analysis revealed changes in body weight in mice and provided model animals for studying the biological functions of Lin28A and Lin28B genes . Methods: Human Lin28A and Lin28B cDNAs were inserted into the downstream of PCAG promoter respectively to construct Lin28A and Lin28B transgene expression vectors. The Lin28A transgenic mice and Lin28B transgenic mice were established respectively by microinjection method. PCR was used to identify the genotypes of transgenic mice, and the mice expressing human Lin28A and Lin28B were screened, and the body weight of the two transgenic mice was counted. Results: (1) The two vectors of Lin28A and Lin28B were successfully constructed by PCR identification and sequencing. ② The genotypes of mice were identified by PCR, and the transgenic mice that could express human Lin28A or Lin28B were screened out. The body weight of transgenic mice and wild-type mice were compared. The body weight of the two transgenic mice was found to be larger than that of the wild-type Mice, indicating that in mice overexpression of Lin28A or Lin28B can cause weight gain in mice. Conclusion: Human Lin28A and Lin28B can express normally in transgenic mice and can exert biological functions. And because Lin28A and Lin28B in mice caused by over-expression can cause weight gain in mice, we speculate that it may affect mice by glucose metabolism caused by weight changes in mice. The constructed transgenic mice of Lin28A and Lin28B provided animal models for further studying the roles of Lin28A and Lin28B in human metabolism, growth and development.