시베리아 철갑상어(Acipenser baerii) 세포주에서 재조합단백질 발현에 관한 연구

Abstract

In spite of many studies about recombinant protein production from variant kinds of organisms, there has been few studies dealing with fish as a host for production of recombinant proteins. Therefore, current status of the researches about recombinant protein production from fish is still primitive suggesting that a fundamental study regarding such a thing should be performed for utilizing fish cells or fish itself as a platform for recombinant protein production. In this regard, here, I evaluated the feasibility of the expression of recombinant proteins in fish cell lines. In order to do that, I tested the effectiveness of lentiviral vector system for gene delivery into the cell lines from Siberian sturgeon (Acipenser baerii) and tried to establish stable cell lines which express human recombinant proteins. Lentiviral vectors containing enhanced green fluorescent protein (EGFP) gene were prepared in host 293LTV cells and treated to 293LTV cells and A. baerii cell lines for delivering EGFP genes within the cells. Lentiviral vector system successfully transferred EGFP genes into 293LTV cells indicating that the system established was operated well. However, it did not transfer EGFP genes into A. baerii cell lines. To evaluate the ability of A. baerii cells to express human recombinant proteins, the expression vectors containing human insulin or erythropoietin (EPO) genes driven by CMV promoter were prepared and transfected into A. baerii cell lines. After transfection, all cell lines transiently expressed insulin or EPO mRNA and subsequently, the cell lines that are consistently expressing insulin or EPO mRNA were established through antibiotic selection and clonal growth. The analyses of insulin or EPO protein expression in each stable cell lines showed that putative insulin or EPO proteins were expressed in the established cell lines suggesting the feasibility of recombinant protein expression in fish cell lines. The results from this study will be able to contribute to development of the system that produces useful recombinant proteins in fish cell lines or fish itself.