人源Plk1及其结构域蛋白的融合表达及纯化

Prokaryotic Expression and Purification of Intact and Domains of Human Plk1

目的:表达纯化GST-Plk1及GST-Plk1 KD、GST-Plk1 PBD融合蛋白,以用于Plk1与其结合蛋白相互作用的研究。方法:PCR扩增Plk1全长及其KD、PBD结构域基因,定向克隆至p GEX-4T-1原核表达载体中,在大肠杆菌中分别表达其融合蛋白,并利用GSH交联的琼脂糖珠纯化。结果:人源Plk1及其结构域基因被克隆至p GEX-4T-1载体中;通过亲和纯化获得带有GST标签的Plk1及其结构域的融合蛋白。结论:构建了GST-Plk1、GST-Plk1 KD、GST-Plk1 PBD表达质粒并表达了相应的融合蛋白,为研究Plk1体外相互作用蛋白提供了基础。

Objective： To express and purify GST-Plk1,GST-Plk1 KD,GST-Plk1 PBD fusion proteins for re-search of interaction between Plk1 and its binding proteins.Methods： The full length of Plk1 gene and its KD,PBD domains genes were amplified by PCR,and then inserted into prokaryotic expression vector p GEX-4T-1.Fu-sion proteins were expressed in Escherichia coli and purified by GSH-agarose beads.Results： Plk1 and its do-mains genes were constructed,and their corresponding GST tag fusion proteins were obtained.Conclusion： The pu-rified fusion proteins will be used in the study of interaction between Plk1 and its binding proteins in vitro.