| abstract |
The present invention provides a method for exogenously expressing BAK1 protein. The method comprises the steps of: digesting the exon gene of the BAK1 extracellular domain with 6 His tags at the C-terminal end with BamH1 and Xho1 enzymes, digesting the pFastBac TM 1 vector with BamH1 and Xho1 enzymes, and ligating the products after the aforementioned enzyme cleavage , construct a vector for exogenous expression of BAK1 protein; the vector is transfected into insect cells, and after culturing the cells, the cells and supernatant are harvested, and the supernatant is purified by chromatography to obtain BAK1 protein with correct conformation, and the expression amount reaches 8.6 mg/ L. The method of the invention overcomes the shortcomings of the prior art that the BAK1 protein in the correct conformation cannot be obtained by using a prokaryotic expression system or the BAK1 protein cannot be expressed by using other eukaryotic expression systems, increases the expression amount of the BAK1 protein with the correct conformation, and saves costs. |