Selection Antibiotics

 

G418 disulfate, hygromycin B, puromycin DiHCl, and blasticidin S HCl are some of the most widely used selection antibiotics in mammalian cell culture media. Selection antibiotics are used as a selective agent after mammalian cells have been transfected with a plasmid containing a gene of interest. To ensure DNA has integrated into the cell's genome, a selective marker is co-transfected with the desired gene which confers resistance to a selection antibiotic or another selective agent. Selective stress is applied to transfected cells by growing transfected cells in the presence of a selection antibiotic and only cells that contain the desired gene along with the selective marker or resistance gene will grow and survive.

 
 

G418 disulfate

Description:
G418 disulfate, also known as G418 sulfate, is routinely used as a selection antibiotic in cell culture gene selection applications. G418 disulfate is an aminoglycoside antibiotic isolated from Micromonospora rhodorangea and is closely related to the gentamicins; most notably, gentamicin B1.

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Mechanism of action:
G418 disulfate, and other aminoglycosides, including kanamycin and neomycin, prevent protein synthesis by blocking the elongation step in prokaryotic and eukaryotic ribosomes.

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Blasticidin S HCl

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.

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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.

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Hygromycin B

Description:
Hygromycin B is a unique aminoglycoside antibiotic derived from Streptomyces hygroscopicus. Hygromycin B is routinely used as a selective agent in cell culture or microbiology applications to isolate hygromycin B resistant cells.

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Mechanism of action:
Hygromycin B inhibits protein synthesis by strengthening the interaction of tRNA binding in the ribosomal A-site. Hygromycin B also prevents mRNA and tRNA translocation by an unknown mechanism. These are unique mechanisms for an aminoglycoside antibiotic and they differ from the mode of action neomycin, gentamicin, and G418.

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Puromycin DiHCl

Description:
Puromycin dihydrochloride is an aminonucleoside antibiotic derived from Streptomyces alboniger. Puromycin dihydrochloride is routinely used as a selective agent in transfection and transformation protocols.

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Mechanism of action:
During translation, puromycin enters the ribosomal “A” site and disrupts peptide transfer. As a result, the ribosome stops and the peptide chain is terminated leading to a nonfunctional protein.

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