FUSION PARTNER LINKAGE AS A STRATEGY FOR OBTAINING A SOLUBLE FORM OF RECOMBINANT HUMAN INTERLEUKIN-4 IN ESCHERICHIA COLI CELLS
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Abstract
The article describes the stages of optimization for the expression of the coding sequence of the human interleukin-4 gene in E. coli cells and its cloning into the pET24b(+) vector under the control of the bacteriophage T7 promoter. Cytokine biosynthesis was induced in E. coli BL21-Gold (DE3) bacterial cells. The target protein was found to accumulate predominantly in the insoluble form. A strategy for interleukin-4 linkage with the fusion partner SUMO is presented. A low biosynthetic potential of E. coli BL21-Gold (DE3) cells carrying fusion constructs was detected, along with the absence of an increase in the yield of the soluble form of IL-4.
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1. Keegan, A. D. Recent advances in understanding the role of IL-4 signaling / A. D. Keegan, W. J. Leonard, J. Zhu // Fac Rev. – 2021. – Vol. 10 – P. 71.
2. IL-4 as a Repurposed Biological Drug for Myocardial Infarction through Augmenta-tion of Reparative Cardiac Macrophages: Proof-of-Concept Data in Mice / Y. Shintani, T. Ito, L. Fields [et al.] // Sci Rep. – 2017. – Vol. 7, nr 1. – P. 68–77.
3. Interleukin-4 Treatment Restores Cellular Immunity After Ethanol Exposure and Burn Injury / K. A. N. Messingham, S. A. Heinrich, E. M. Schilling, E. J. Kovacs // Alcoholism Clin Exp Res. – 2002. – Vol. 26, nr 4. – P. 519–526.
4. Fabrication of Interleukin-4 Encapsulated Bioactive Microdroplets for Regulating In-flammation and Promoting Osteogenesis / Y. Zhang, J. Cao, M. Jian [et al.] // IJN. – 2023. – Vol. 18 – P. 2019–2035.
5. Advanced therapy medicinal products: current and future perspectives / E. Hanna, C. Rémuzat, P. Auquier, M. Toumi // J Mark Access Health Policy. – 2016. – Vol. 4. – P. 103–113.
6. Schmidt, F. R. Recombinant expression systems in the pharmaceutical industry / F. R. Schmidt // Appl Microbiol Biotechnol. – 2004. – Vol. 65, nr 4. – P. 363–372.
7. Jia, B. High-throughput recombinant protein expression in Escherichia coli : current status and future perspectives / B. Jia, C. O. Jeon // Open Biol. – 2016. – Vol. 6, nr 8. – P. 160–196.
8. Overton, T. W. Recombinant protein production in bacterial hosts / T. W. Overton // Drug Discovery Today. – 2014. – Vol. 19, nr 5. – P. 590–601.
9. Baneyx, F. Recombinant protein folding and misfolding in Escherichia coli / F. Baneyx, M. Mujacic // Nat Biotechnol. – 2004. – Vol. 22, nr 11. – P. 1399–1408.
10. Brondyk, W. H. Chapter 11 Selecting an Appropriate Method for Expressing a Re-combinant Protein / W. H. Brondyk // Methods in Enzymology // Elsevier, 2009. – Vol. 463. – P. 131–147.
11. SUMO fusion technology for difficult-to-express proteins / T. R. Butt, S. C. Edavet-tal, J. P. Hall, M. R. Mattern // Protein Expr Purif. – 2005. – Vol. 43, nr 1. – P. 1–9.
12. Comparison of SUMO fusion technology with traditional gene fusion systems: Enhanced expression and solubility with SUMO / J. G. Marblestone, S. C. Edavettal, Y. Lim [et al.] // Protein Sci. – 2006. – Vol. 15, nr 1. – P. 182–189.
13. Current protocols in molecular biology // ed. by F. M. Ausubel. – New York : Wiley, 1987. – Vol. 1.
14. Sanger, F. DNA sequencing with chain-terminating inhibitors / F. Sanger, S. Nicklen, A. R. Coulson // Proc Natl Acad Sci U S A. – 1977. – Vol. 74, nr 12. – P. 5463–5467.
15. Laemmli, U. K. Cleavage of structural proteins during the assembly the head of bac-teriophage T4 / U. K. Laemmli // Nature. – 1970. – Vol. 227. – P. 680–685.
16. Protein Identification and Analysis Tools on the ExPASy Server / E. Gasteiger, C. Hoog¬land, A. Gattiker [et al.] // The Proteomics Protocols Handbook. – New Jersey : Humana Press, 2005. – P. 571–607.
17. Burland, T. G. DNASTAR’s Lasergene Sequence Analysis Software / T. G. Burland // Bioinformatics Methods and Protocols. – New Jersey : Humana Press, 1999. – Vol. 132. – P. 71–91.
18. Quantifying Western blots: Pitfalls of densitometry / M. Gassmann, B. Grenacher, B. Rohde, J. Vogel // Electrophoresis. – 2009. – Vol. 30, nr 11. – P. 1845–1855.
19. Schneider, C. A. NIH Image to ImageJ: 25 years of Image Analysis / C. A. Schnei-der, W. S. Rasband, K. W. Eliceiri // Nat Methods. – 2012. – Vol. 9, nr 7. – P. 671–675.
20. Expression of recombinant human IL-4 in Pichia pastoris and relationship between its glycosylation and biological activity / R. Li, C. Xie, Y. Zhang [et al.] // Protein Expression and Purification. – 2014. – Vol. 96. – P. 1–7.
21. Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system / A. Munir, N. Ahmed, M. Akram [et al.] // Biotechnol Lett. – 2023. – Vol. 45, nr 8. – P. 1001–1011.