• Blasticidin S HCl packaged and labeled in glass bottle.
  • Molecular structure of blasticidin S HCl.

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SKU: B001

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Description

Blasticidin S HCl is a nucleoside antibiotic derived from Streptomyces griseochromogenes. Blasticidin S HCl is routinely used in cell culture gene selection applications as a selectable agent to isolate blasticidin S resistant mammalian and bacterial cells.

TOKU-E recommends preparing stock solutions at 5-10 mg/mL in water or 20mM HEPES. Stock solutions can be stored at 4°C for short term storage and -20°C for long term storage.

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Lu et al. used blasticidin S HCl and puromycin dihydrochloride from TOKU-E to select for transfected AS-B145 and BT-474 cells. "Ovatodiolide Inhibits Breast Cancer Stem/Progenitor Cells through SMURF2-Mediated Downregulation of Hsp27"  

    CAS Number

    3513-03-9

    Molecular Formula

    C17H26N8O5 · HCl

    Molecular Weight

    458.90

    Mechanism of Action

    Blasticidin S HCl 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. 

    Storage Conditions

    2-8°C

    Tariff Code

    2934.99.9000

    Spectrum

    Blasticidin S HCl is biologically active against susceptible mammalian and prokaryotic cells.

Applications

    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.

    Microbiology Applications

    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.

Specifications

    Form

    Powder

    Appearance

    White or off-white powder

    Source

    Streptomyces griseochromogenes

    Potency (on a dry basis)

    ≥850µg/mg

    Endotoxin

    ≤1.0EU/mg

References

    References

    Yamaguchi, I., et al. "Expression of the Blasticidin S Deaminase Gene (bsr) in Tobacco: Fungicide Tolerance and a New Selective Marker for Transgenic Plants." Molecular and General Genetics 223.2 (1990): 332-34. www.ncbi.gov. Web. 7 Sept. 2012.

    "Blasticidin S Inhibits Translation by Trapping Deformed TRNA on the Ribosome." PNAS 110.30 (2013): 12283-2288. Ncbi.nlm.nih.gov. Web. 4 Sept. 2014.

    Bento, Fernanda M. "Over Expression of the Selectable Marker Blasticidin S Deaminase Gene Is Toxic to Human Keratinocytes and Murine BALB/MK Cells." BMC Biotechnology 4.29 (2004): 1-10. Springer.com. BioMed Central. Web. 4 Nov. 2014.

    Kimura, M. "Blasticidin S Deaminase Gene from Aspergillus Terreus (BSD): A New Drug Resistance Gene for Transfection of Mammalian Cells." Biochemica Et Biophysica Acta. 1219.3 (1994): 653-59. Ncbi.nlm.nih.gov. Web. 4 Nov. 2014.

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