Blasticidin S deaminase gene from Aspergillus terreus (BSD): a new drug resistance gene for transfection of mammalian cells.
Blasticidin S deaminase (BSD) is a drug inactivating enzyme produced by Aspergillus terreus, which convert blasticidin S (BS) to a non-toxic deamino-hydroxy derivative. The BSD gene was fused to SV 40 transcriptional regulatory elements and the resulting vector was used to transfect FM3A cells. Expression of BSD conferred resistance to BS and allowed efficient isolation of integrative transfectants which have stably maintained the BS-resistance phenotype after repeated transfer to fresh selective medium. The frequency of transfection was comparable to that with neo and about 80-times greater than with bsr, a BS-resistance gene of bacterial origin which can be used to isolate efficiently transfectant HeLa cells. Using BSD as a selectable marker, we obtained several stable cell lines expressing the firefly luciferase gene. Four independent transfectants among the randomly selected 5 BS-resistance colonies exhibited detectable luciferase activity under the control of dexamethasone-inducible promoter in the expression vector. The successful application of BSD strongly suggests the usefulness of BS as a versatile selective reagent for introduction of cloned DNA sequences into mammalian cells.
Kimura
M
M
Microbial Toxicology Laboratory, Institute of Physical and Chemical Research (RIKEN) Saitama, Japan.
Takatsuki
A
A
Yamaguchi
I
I
D000619:Aminohydrolases / Q000235:genetics*; D000818:Animals; D001230:Aspergillus / Q000235:genetics*; D001483:Base Sequence; D001517:Coleoptera / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009705:Nucleosides / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D012641:Selection, Genetic; D014162:Transfection
D005819:Genetic Markers; D009705:Nucleosides; C004500:blasticidin S; D008156:Luciferases; D000619:Aminohydrolases; C019795:blasticidin S deaminase