猪FoxN1基因转录变异体亚细胞定位研究

Research on the subcellular localization of pig transcriptional variants of FoxN1 gene

Fox N1基因主要表达于胸腺上皮细胞和皮肤角质细胞,在胸腺发育、T细胞增殖、毛发生长、指甲发育等方面发挥重要作用。Fox N1基因存在多种转录变异体,人类Fox N1基因已发现18种转录变异体。转录变异体的存在使基因差异表达和蛋白质多样性存在可能。为深入研究转录变异体生物学功能,研究将前期发现的猪Fox N1基因3种转录变异体分别连入p EGFP-N1载体内,构建各自融合蛋白,转入猪PK15细胞,利用荧光显微镜观察转录变异体亚细胞定位情况。结果表明,p EGFP-N1空载体转染表达绿色荧光蛋白在PK15细胞核和细胞质均匀分布,而重组载体p EGFP-N1-Fox N1(a/b/c)表达的融合蛋白仅在细胞核内均匀分布。结果可为3种转录变异体后续功能研究奠定基础。

Fox N1 gene was mainly expressed in thymic epithelial cell and skin keratinocytes. It played important roles in thymus development, T cell proliferation, hair growth and nail development. There were many transcriptional variants in the Fox N1 gene. It had been found 18 transcriptional variants in human Fox N1 gene. The presence of transcriptional variants made it possible of gene differential expression and protein diversity. In previous study, three transcriptional variants of pig Fox N1 gene were found. In order to deep research on the functional of every transcriptional variant, three transcriptional variants of pig Fox N1 gene were connected with p EGFP-N1 vector. Fusion protein of everyone was constructed and transfected in PK15 cell. The results showed that, only EGFP expression could be detected in PK15 cell nucleus and cytoplasm. Three fusion proteins only could be detected in cell nucleus. This result could prove the basic knowledge for further research on pig Fox N1 gene.