Article Sidebar

PDF
Date Published: 27/06/2023
Abstract Views: 123
Views PDF: 109
DOI: https://doi.org/10.51298/vmj.v527i1B.5741
Issue
Vol. 527 No. 1B (2023)
Section
Các bài báo
How to Cite
Nguyễn, Q. T., Nguyễn, L. T. H., Mai, H. N., & Thái, K. M. (2023). EXPRESSION OF HUMAN INTERLEUKIN-2 FUSED WITH SUMO IN ESCHERICHIA COLI. Vietnam Medical Journal, 527(1B). https://doi.org/10.51298/vmj.v527i1B.5741

EXPRESSION OF HUMAN INTERLEUKIN-2 FUSED WITH SUMO IN ESCHERICHIA COLI

Quốc Thái Nguyễn1,, Lê Thanh Huyền Nguyễn1, Huỳnh Như Mai1, Khắc Minh Thái1
1 UMP

Main Article Content

Abstract

Background: Interleukin-2 (IL-2) is used as a therapeutic agent in the treatment of cancer and autoimmune diseases. However, in Vietnam, the price of IL-2 used for in vitro experiments is expensive. Objectives: This study aims to express recombinant human IL-2 fused with SUMO in Escherichia coli. Methods: Plasmid pET-SUMO-il2 was transformed into E. coli BL21(DE3) for the expression of IL-2 with SUMO tag. Expression conditions for optimal soluble recombinant IL-2 were examined, including culture medium (LB, TB, ZYP-5052); IPTG concentration (0.02, 0.1, and 0.5 mM); culture temperature (25°C and 37℃). The recombinant IL-2 fused with SUMO tag was purified by immobilized metal affinity chromatography method (IMAC). Results: The transformed bacteria can express SUMO-IL-2 with high yield in TB medium induced by 0.1 mM IPTG at 25°C. From 200 mL of bacterial culture, we obtained approximately 108 mg of purified protein by IMAC. Conclusions: In this study, we have successfully transformed E. coli BL21(DE3) with the recombinant plasmid pET-SUMO-il2. The bacteria could express soluble SUMO-IL-2 with high yield and the pure protein was obtained by 1-step purification using Ni Sepharose column.    

Article Details

Keywords

: recombinant human interleukin-2, pET-SUMO, E. coli

References

1. Abbas, A. K., Trotta, E., R. Simeonov, D., Marson, A., & Bluestone, J. A. (2018). Revisiting IL-2: Biology and therapeutic prospects. Science immunology, 3(25):1482.
2. Nguyễn Quốc Thái, Nguyễn Lê Thanh Huyền, Mai Huỳnh Như, Thái Khắc Minh (2023), “Dự đoán khả năng hoà tan và thiết kế vector biểu hiện interleukin-2 người trên Escherichia coli”, Tạp chí Y học Việt Nam, Tập 526, tháng 6, số X, tr.
3. Studier, F. W. (2005) “Protein production by auto-induction in high-density shaking cultures”, Protein expression and purification, 41(1), pp. 207-234.
4. Bradford, M. M. (1976). “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding”, Analytical biochemistry, 72(1-2), pp. 248-254.
5. Panavas, T., Sanders, C., & Butt, T. R. (2009). SUMO fusion technology for enhanced protein production in prokaryotic and eukaryotic expression systems. SUMO protocols, 303-317.
6. Zhang, M., Jiang, X., Su, Z., Lin, J., Xiang, Q., Yang, Z.,... & Li, X. (2012). Large-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli. Applied microbiology and biotechnology, 93, 613-621.
7. Peciak, K., Tommasi, R., Choi, J. W., Brocchini, S., & Laurine, E. (2014). Expression of soluble and active interferon consensus in SUMO fusion expression system in E. coli. Protein expression and purification, 99, 18-26.
8. Tileva, M., Krachmarova, E., Ivanov, I., Maskos, K., & Nacheva, G. (2016). Production of aggregation prone human interferon gamma and its mutant in highly soluble and biologically active form by SUMO fusion technology. Protein Expression and Purification, 117, 26-34.