人Polo样激酶1基因的克隆表达及其影响c-Abl表达水平功能的初探

Gene Cloning and Expression of Human Plk1 and Exploring Study of its Effect on c-Abl Expression

目的:克隆并在293细胞中表达人Polo样激酶1(Plk1)基因,探索Plk1对非受体型酪氨酸激酶c-Abl表达水平的影响。方法:利用PCR法扩增Plk1基因,分别定向克隆至pcDNA3-Flag及pCMV-Myc真核表达载体,将上述质粒分别转染293细胞进行瞬时表达,Western印迹检测Plk1蛋白的表达;将上述质粒分别与c-Abl表达质粒共转293细胞,检测带有不同标签的Flag-Plk1或Myc-Plk1对细胞中c-Abl激酶表达的影响。结果:构建了Flag-Plk1和Myc-Plk1真核表达质粒,其在293细胞中均可表达,蛋白的相对分子质量为68×103;与其共转的c-Abl激酶表达水平显著下调。结论:在293细胞中表达了Flag-Plk1和Myc-Plk1蛋白,且初步发现Plk1可以抑制c-Abl的表达。

Objective： To clone and express human Polo-like kinase-l（Plkl） gene in 293 cell lines, and study its effect on the expression of non-receptor tyrosine kinase c-Abl. Methods： Plkl gene was amplified by PCR, and then inserted into eukaryotic expression vector pcDNA3-Flag and pCMV-Mye respectively. Those recombinant expression plasmids were transiently transfected into 293 cell lines, and the expression of Plkl was identified by Western blotting. Different tagged Plkl and c-Abl were cotransfected into 293 cells to identify the role of Plkl on the expression of c-Abl kinase. Results： The Flag-Plkl and Myc-Plkl eukaryotic expression plasmids were constructed, and expressed in 293 cell lines with the molecular weight of 68 kD. The expression of c-Abl reduced significantly when contrandfected with tagged Plkl or c-Abl plasmids into 293 cells. Conclusion： Flag-Plkl and Myc-Plkl have been expressed in 293 cells, and it was found that Plkl can inhibit expression of c-Abl.