What does neomycin phosphotransferase do?
What does neomycin phosphotransferase do?
Neomycin phosphotransferase II confers resistance to kanamycin and neomycin in bacteria and G418 (Geneticin, G418 Sulfate) in mammalian cells. These antibiotics bind to ribosomal components and inhibit protein synthesis.
What is neomycin resistance gene?
The neo (neomycin-resistance) gene of transposon Tn5 encodes the enzyme neomycin phosphotransferase II (EC 2.7. 1.95), which confers resistance to various aminoglycoside antibiotics, including kanamycin and G418.
How long does hygromycin selection take?
Puromycin usually kills cells in 3 days, G418 in 7 days, hygromycin in 7 days and blasticidin in 5 days. Cells are often split before selection (e.g. 1:3). That’s because selection does not work if cells are too confluent! Cells should not be too sparse either; they would have a difficult time recovering.
What is a degradation product of the aminoglycoside Neomycin?
Neamine, an Antibacterial Degradation Product of Neomycin.
Is hygromycin temperature sensitive?
– Hygromycin B is stable for three months at room temperature, two years at 4°C, and two years at -20°C. Avoid repeated freeze-thaw cycles. – Hygromycin B is sensitive to high concentrations of acid but a short-term exposure to dilute acids can be tolerated.
How long do hygromycin plates last?
The antibiotic is stable for at least 2 years at 4°C. It is stable for about one month at 37°C.
What is puromycin nacetyl transferase?
Puromycin is used to select for cell lines that have been transformed by vectors that express puromycin-N-acetyl-transferase (pac), e.g., pPUR (Cat. No. 631601). (3′-[α-Amino-p-methoxy-hydrocinnamamido]-3′-deoxy-N, N-dimethyladenosine) dihydrochloride. This is the 25 mg format.
What is the recommended concentration for G418 and puromycin?
G418, Puromycin, & Hygromycin B Recommended concentration (µg/ml) Recommended concentration (µg/ml) Antibiotic Selection Maintenance G418 100–800 200 Puromycin 0.25–10 0.25 Hygromycin B 50–400 100
Is G418 sulfate the same as hygromycin?
G418 sulfate: An aminoglycoside antibiotic similar to gentamycin. Toxic to bacteria, yeast, higher plants, and mammalian cells, in addition to protozoans and helminths Hygromycin B: An aminoglycoside antibiotic that inhibits protein synthesis in bacteria, fungi, and higher eukaryotes
What is hygromycin B used for?
Hygromycin B: An aminoglycoside antibiotic that inhibits protein synthesis in bacteria, fungi, and higher eukaryotes Puromycin dihydrochloride: A broad spectrum antibiotic that inhibits protein synthesis in both prokaryotic and eukaryotic organisms.
