活化型TAK1转基因小鼠的建立及TAK1对牙釉质发育影响的初步研究

发布时间：2018-02-25 16:18

本文关键词： TAK 基因突变转基因动物釉质发育　出处：《牙体牙髓牙周病学杂志》2014年03期 　论文类型：期刊论文

【摘要】：目的:建立人活化型TAK1(caTAK1)转基因小鼠,初步研究TAK1对釉质发育的影响。方法:克隆小鼠釉原蛋白(Amelogenin,Amelx)启动子及人TAK1的全长基因,并利用缺失突变原理扩增caTAK1基因;运用Gateway技术将caTAK1基因插入Amelx启动子下游构建转基因表达载体,并通过显微注射法建立caTAK1转基因小鼠;用PCR扩增对转基因小鼠的基因型进行鉴定,RT-PCR检测外源性TAK1基因在转基因小鼠颌骨中的表达,并运用X线片、体视显微镜及微型CT对转基因小鼠磨牙釉质的矿化程度和病理性磨损程度进行观察。结果:成功构建了caTAK1转基因表达载体并制备转基因小鼠;转入的人caTAK1基因在小鼠颌骨组织中特异性表达;活化型TAK1转基因小鼠磨牙釉质的矿化程度较野生型小鼠差,1月龄时,两种小鼠磨牙釉质表面的磨损程度无明显差别,而6月龄时,与野生型小鼠相比,活化型TAK1转基因小鼠磨牙釉质有严重磨损。结论:TAK1在牙齿釉质发育过程中具有重要作用。
[Abstract]:Objective: to establish human activated TAK1 caTAK1) transgenic mice and to study the effect of TAK1 on the development of enamel. Methods: the full-length gene of mouse amelogenin Amelx promoter and human TAK1 were cloned, and the caTAK1 gene was amplified by the principle of deletion mutation. The caTAK1 gene was inserted into the downstream of the Amelx promoter by Gateway technique to construct the transgenic expression vector, and the caTAK1 transgenic mice were established by microinjection. The genotypes of transgenic mice were identified by PCR amplification. The expression of exogenous TAK1 gene in the jaws of transgenic mice was detected by RT-PCR. The mineralized degree and pathological wear degree of grinding enamel in transgenic mice were observed by stereoscopic microscope and microCT. Results: caTAK1 transgenic expression vector was successfully constructed and transgenic mice were prepared. The specific expression of human caTAK1 gene was found in the mandibular tissue of mice, and there was no significant difference in the wear degree of the enamel surface between the two mice at the age of 1 month compared with the wild-type mice, and the mineralization degree of the grinding enamel in the activated TAK1 transgenic mice was less than that in the wild-type mice. Compared with wild-type mice, activated TAK1 transgenic mice showed severe wear in grinding enamel at 6 months old. Conclusion: Tak 1 plays an important role in the development of tooth enamel.
【作者单位】：滨州医学院附属医院口腔科;
【基金】：国家自然科学基金资助项目(81170927)
【分类号】：R780.2

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