• Blasticidin S HCl, 10 mg/mL packaged and labeled.

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SKU: B006-B007

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Description

Blasticidin S HCl is a peptidyl nucleoside produced by several species of Streptomyces that was first isolated from S. griseochromogenes in 1958.  Blasticidin S inhibits protein synthesis and is active against bacteria, fungi, nematodes, and tumor cells.  The compound is used as a selection antibiotic for both eukaryotic and prokaryotic cells, and a marker for strain manipulation.

 TOKU-E carries two forms of Blasticidin S HCl:

 Blasticidin S HCl solution contains 10 mg/mL blasticidin S HCl in 20mM HEPES.

B006 (10 x 1 mL) contains 10 mg blasticidin S HCl per vial (100 mg total).

B007 (20 mL) contains 200 mg blasticidin S HCl per vial.

This product is considered a dangerous good. Quantities above 1 g may be subject to additional shipping fees.

    CAS Number

    3513-03-9

    Molecular Formula

    C17H26N8O5 · HCl

    Molecular Weight

    458.90 g/mol

    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

    -20°C

    Tariff Code

    2934.99.9000

Applications

    Eukaryotic Cell Culture Applications

    Blasticidin is a selection antibiotic for both eukaryotic and prokaryotic cells. Resistance to Blasticidin is conferred by the following genes:

    1) bsr (blasticidin resistance), from Bacillus aureus>/em>, which codes for blasticidin-S deaminase.

    2) bls (blasticidin S acetyltransferase), from Streptoverticillum spp.

    3) BSD (blasticidin S deaminase), from Apergillus terreus.

    Researchers used Blasticin S from TOKU-E to select for transfected AS-B145 and BT-474 cells, which are human breast cancer stem/progenitor cells, a subpopulation of cancer cells that are involved in tumor initiation, resistance to therapy, and metastasis (Lu et al, 2016). Typically, mammalian cells are sensitive to Blasticidin S HCl concentrations of 1-10 µg/ml, and bacteria to 25-100 µg/ml. Optimal selection concentration depends on the cell line, growth conditions, media, reagent quality and potency, manufacturing lot, cell density, cell metabolic rate, cell cycle phase, and plasmid properties. A kill curve should be performed for each experimental system to determine the optimal working concentration.

    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

    Solution (sterile)

    Appearance

    Clear and colorless or light yellow solution

    Source

    Streptomyces griseochromogenes

    pH

    7.2-7.5

    Concentration

    10.0±0.3 mg/mL

References

    References

    Adachi H, Hasebe T, Yoshinaga K , Ohta T and Sutoh K (1994) Isolation of Dictyostelium discoideum cytokinesis mutants by restriction enzyme-mediated integration of the Blasticidin S resistance marker. Biochem. Biophys. Res. Comm. 205(3):1808-1814

    Bento, FM (2004) Over Expression of the Selectable Marker Blasticidin S Deaminase Gene Is Toxic to Human Keratinocytes and Murine BALB/MK Cells." BMC Biotechnol. 4 (29):1-10 PMID 15575952.

    Izumi M. et al., 1991. Blasticidin S-resistance gene (bsr): A novel selectable marker for mammalian cells. Exp.Cell Res.197:229-33

    Lu K-T et al (2016) Ovatodiolide inhibits breast cancer stem/progenitor cells through SMURF2-mediated downregulation of Hsp27. Toxins 8(5):127.

    Kimura M, Takatsuki A, Yamaguchi I (1994) Blasticidin S deaminase gene from Aspergillus terreus(BSD): A new drug resistance gene for transfection of mammalian cells. Biochim. Biophys. Acta. 1219(3):653-65 PMID 7948022

    Svidritskiy E, Ling C, Ermolenko DN, Korostelev AA (2013) Blasticidin S Inhibits Translation by Trapping Deformed TRNA on the Ribosome. PNAS 110(30):12283-12288 PMID 23824292

    Takeuchi S, Hirayama K, Ueda K, Sakai H and Yonehara H (1958) Blasticidin S, a new antibiotic. J. Antibiot. 11(1):1-5 PMID 13525246

    Yamaguchi I et al (1990) Expression of the Blasticidin S Deaminase Gene (bsr) in Tobacco: Fungicide Tolerance and a New Selective Marker for Transgenic Plants. Mol. Gen. Genet (2):332-334 PMID 2250657

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