Pin1在皮肤组织中的表达以及可诱导转基因小鼠模型的建立

摘要

Objective To observe the Pin1 expression pattern in skin and to establish an inducible skin specific Pin1 overexpression mouse model. Methods The mouse Pin1 gene was cloned into modified vector pTRE2 with C⁃terminal Myc tag. The linearized pTRE2⁃Pin1 DNA was micro⁃injected into one⁃cell embryos followed by implantation into foster mice to produce TRE⁃Pin1 transgenic mice. Results TRE⁃Pin1 transgenic founder mice were successfully created. These mice were crossed with transgenic tool mice K14⁃rtTA to create epithelial specific double transgenic progenies. Pin1 gene was induced by incorporating doxycycline into drinking water of the mice. Pin1 protein overexpression in the skin was con⁃firmed by Western blot and immunohistochemistry. The endogenous Pin1 protein was predominantly expressed in epidermal cells in the skin. Conclusions The inducible skin specific Pin1 overexpression mouse model is successfully established which may serve as a useful model for further study of Pin1 functions in the skin.%目的：观察Pin1在皮肤中的表达情况，构建Pin1在皮肤中可诱导表达的转基因小鼠模型。方法将小鼠Pin1基因克隆到改造过的可与Myc标签蛋白融合的pTRE2载体中，并将线性化的DNA通过显微注射的方式构建TRE⁃Pin1小鼠。结果成功获得TRE⁃Pin1转基因首建鼠，该小鼠与上皮特异的K14⁃rtTA转基因小鼠配繁，获得Pin1在皮肤上皮特异性可诱导表达的双转基因鼠；通过将多西霉素（又名强力霉素，Doxycycline）加入饮水的方式诱导Pin1基因的表达，并通过Western blot，免疫组织化学等方式证明了Pin1蛋白在皮肤上皮中能特异性地过表达。我们还发现内源的Pin1在皮肤中主要表达于上皮细胞。结论成功构建了Pin1在皮肤中可诱导表达的转基因小鼠模型，为后续研究Pin1在皮肤中的功能奠定基础。