CONDITIONS OF SELECTION

Escherichia coli :

Blasticidin S resistant transformants can be selected in low salt LB agar medium, pH 8, supplemented with 100 µg/ml blasticidin S. For optimum results the use of Fast-Media® BLAS is recommended.

Mammalian cells :

The working concentrations of blasticidin S for mammalian cell lines varie from 3 to 50 µg/ml. In a starting experiment we recommend to determine optimal concentrations of antibiotic required to kill your host cell line. After treatment, cell death occurs rapidly allowing the selection of transfected cells with plasmids carrying the BSD or bsr genes in as little as 7 days post-transfection. Suggested working conditions for selection in some mammalian cells are listed below:

Cell line
Species
Tissue
Culture medium
Blasticidin (µg/ml)
293
Human
Kidney
DMEM
3-10
HeLa
Human
Uterus
DMEM
3-10
B16
Mouse
Melanoma
RPMI
3-10
PC1.0
Hamster
Pancreatic adenocarcinoma
RPMI
10-30

References

  • IZUMI M., MIYAZAWA H., KAMAKURA T.,YAMAGUCHI I., ENDO T.and F.HANAOKA. 1991. Blasticidin S-resistance gene (bsr): a novel selectable marker for mammalian cells.Exp.Cell Res.197: 229-233
  • KIMURA M.,TAKATSUKI A and I.YAMAGUCHI. 1994. Blasticidin S deaminase gene from Aspergillus terreus (BSD) a new drug resistance gene for transfection of mammalian cells. Biochim.Biophys.Acta.1219:653-659