Description Blasticidin S is a nucleoside produced by several species of Streptomyces, first reported in the late 1950s. Blasticidin S is an antifungal agent with particularly potent activity against the rice pathogen, Piricularia oryzae, for which it was used commercially for some time in Japan. Blasticidin S inhibits protein synthesis and is active against bacteria, tumor cell lines and nematodes. More recently, blasticidin S has been used as a marker for strain manipulations. Blasticidin S should be used as the free base to avoid problems associated with the use of the hydrochloride.
Blasticidin S is soluble in ethanol, methanol, DMF and DMSO.
Mechanism of Action
Blasticidin S inhibits protein synthesis in prokaryotic and eukaryotic cells by binding to the ribosomal P-site which strengthens tRNA binding and slows down and prevents subsequent peptide synthesis.
Mechanisms of resistance
Resistance to blasticidin S is conferred by bsr, BSD, and bls resistance genes isolated from Bacillus cereus K55-S1, Aspergillus terreus, and Streptoverticillum spp, respectively.
The bsr resistance gene is a 420 bp fragment and encodes a 15 kDa blasticidin S deaminase which catalyzes the reaction of blasticidin S to deaminohydroxyblasticidin S. Deaminohydroxyblasticidin S is a biologically inactive derivative of blasticidin S and does not interact with or inhibit prokaryotic or eukaryotic ribosomes.
The bsd resistance gene is a 393 bp fragment and also encodes a blasticidin S deaminase enzyme which catalyzes a similar reaction to the BSR deaminase. A study by Kimura et al. found the transfection frequency with bsd to be 80X greater than with bsr when using FM3A cells.
The bls gene resistance gene encodes an acetyltransferase which interacts with acetyl-coenzyme A and prevents blasticidin S from inhibiting protein synthesis.
Blasticidin S HCl is biologically active against susceptible mammalian and prokaryotic cells.
Blasticidin S HCl can be used as a selection agent after transformation of prokaryotic (bacterial) cells, namely E. coli. Optimal blasticidin S HCl selection concentrations range from 25 - 100 µg/mL and should be tested for each experimental condition. Selective media containing blasticidin S HCl should contain a low salt concentration (<90mM) and pH ≤7 to avoid blasticidin degradation.
Eukaryotic Cell Culture Applications
Blasticidin S HCl is frequently used as a selection agent after transfection of eukaryotic cells. Selectable markers include blasticidin S deaminase resistant genes bsr and BSD. The optimal selection concentration of blasticidin S HCl resistant mammalian clones depends on the cell lines used, growth conditions, media, qualtiy and potency of blasticidin S HCl, cell density, cell metabolic rate, cell cycle phase, and the plasmid carrying the blasticidin resistance genes. A kill curve should therefore be performed for every experimental system and for every lot of blasticidin S HCl to determine the optimal working concentration for the selection of blasticidin S resistant cells. Optimal selection concentrations of blasticidin S range from 1 µg/mL - 30 µg/mL but typically lie between 2 µg/mL and 10 µg/mL.
For more information on relevant cell lines, culture medium, and working concentrations, please visit the TOKU-E Cell-culture Database.
Blasticidin S, a new antibiotic. Takeuchi S. et al. J. Antibiot. 1958, 11, 1.
Nucleoside antibiotics. Suhadolnik R.J. pub. Wily-Interscience. 1970, 298.
Blasticidin S-resistance gene (bsr): a novel selectable marker for mammalian cells. Izumi M. et al. Exp. Cell Res. 1991, 197, 229.
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