Overexpression, isolation and purification of functionally active bacterioferritin Dps E.coli
Keywords:
Escherichia coli, bakterioferritin Dps, DNA, chromatography.
Abstract
A method for isolation and purification of the recombinant Dps protein from Escherichia coli
was suggested. The procedure involves cloning of the dps gene into the pGEM vector, followed by the
induction of the recombinant gene expression within BL21*(DE3) E. coli cells, ion-exchange
chromatography on DEAE-Sephadex-A25 (medium), protein fractionation with ammonium sulfate and
gel filtration on Sephadex-G200. The developed method allows to obtain electrophoretically
homogeneous and functionally active Dps protein
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References
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R., Artymiuk P. J., Harrison P. M. Insights into the effects on metal binding of the
systematic substitution of five key glutamate ligands in the ferritin of Escherichia coli // J.
Biol. Chem. 2003. V. 278. P. 26275-26286.
2. Zhao G., Ceci P., Ilari A. et al. Iron and hydrogen peroxide detoxification properties
of DNA-binding protein from starved cells // J. Biol. Chem. 2002. V. 277. P. 27689–
27696.
3. Haikarainen T., Papageorgiou A.C. Dps-like proteins: structural and functional
insights into a versatile protein family // Cell Mol. Life Sci. 2010. V. 67. P. 341–351.
4. Grant R.A., Filman D.J., Finkel S.E. et al. The crystal structure of Dps, a ferritin
homolog that binds and protects DNA // Nat. Struct. Biol. 1998. V. 5(4). P. 294–303.
5. Антипов С. С., Курганов К. В., Чемерис К. С. и др. Бактериоферритин как
биосенсор и наноструктура // Вестник биотехнологии и физико-химической
биологии им. Ю. А.Овчинникова. 2007. Т. 3. С. 40-44.
6. Швырева У.С., Тутукина М. Н., Озолинь О.Н. Бактериоферритин: свойства и
структурно-функциональная организация регуляторной области гена Dps //
Биофизика. 2011. Т. 56(2). С. 821-830
7. Ali Azam T., Iwata A., Nishimura A. et al. Growth phase-dependent variation in
protein composition of the Escherichia coli nucleoid // J. Bacteriol. 1999. V. 181. P. 6361–
6370.
8. Davis B.J. Disc electroforesis II. Method and application to human serum protein //
Ann. N.Y. Acad. Sci. 1994. Vol. 121. P. 404-427.
9. Shavkunov K. S., Masulis I. S., Tutukina M. N. et al. Gains and unexpected lessons
from genome-scale promoter mapping // Nucleic Acids Res. 2009. V. 37. P. 4919-4931.
10. Shevchenko A., Wilm M., Vorm O. Mass spectrometric sequencing of protein from
silver-stained polyarylamide gels // Anal. Chem., 1996. V.68. P 850 – 858.
11. Almiron M., Link A.J., Furlong D. et al. A novel DNA-binding protein with
regulatory andprotective roles in starved Escherichia coli // Genes Dev. 1992. V. 6. P.
2646–2654.
12. Azam T. A., Ishihama A. Twelve Species of the Nucleoid-associated Protein from
Escherichia coli. Sequence recognition specificity and DNA binding affinity // J. Biol.
Chem., 1999. V. 274. P. 33105-33113.
13. Ceci P., Cellai S., Falvo E. et al. DNA condensation and self-aggregation of
Escherichia coli Dps are coupled phenomena related to the properties of the N-terminus //
Nucleic Acids Res. 2004. V. 32. P. 5935–5944
Published
2019-11-25
How to Cite
Pokusaeva, V. O., Antipov, S. S., Shvyreva, U. S., & Tutukina, M. N. (2019). Overexpression, isolation and purification of functionally active bacterioferritin Dps E.coli. Sorbtsionnye I Khromatograficheskie Protsessy, 12(6). Retrieved from https://journals.vsu.ru/sorpchrom/article/view/1891
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