What is hygromycin phosphotransferase?
The Escherichia coli hygromycin phosphotransferase (hph) gene, which confers hygromycin resistance, is commonly used as a dominant selectable marker in genetically modified bacteria, fungi, plants, insects, and mammalian cells.
What is hygromycin gene?
The resistance gene is a kinase that inactivates hygromycin B through phosphorylation. Since the discovery of hygromycin-resistance genes, hygromycin B has become a standard selection antibiotic in gene transfer experiments in many prokaryotic and eukaryotic cells.
What does hygromycin B do?
Hygromycin B is an aminoglycosidic antibiotic produced by Streptomy- ces hygroscopicus that kills bacteria, fungi, and higher eukaryotic cells by inhibiting protein synthesis. It has been reported to interfere with translocation (1, 2) and to cause mistranslation (3).
What concentration of hygromycin is used for selection?
100-1000 µg/ml
The concentration for selection ranges from 100-1000 µg/ml (typically 200 µg/ml) and should be optimized for each cell line.
How does hygromycin selection work?
Specifically, it inhibits protein synthesis by interfering with translocation of the 70S ribosome subunit and inducing misreading of the mRNA template. Hygromycin B has been used to select transfectants in a wide variety of cells including bacteria, protozoans, yeast, fungi, plants, and mammalian cells.
How do you make a hygromycin B stock solution?
Preparation Instructions Hygromycin B is soluble in water at >50 mg/mL. 2 It is also soluble in methanol or ethanol. Solutions should be sterilized by filtration, not by autoclaving. Hygromycin B is dissolved in water at 50 mg/mL, then sterile-filtered to produce H 0654, or γ-irradiated to produce H 5527.
What is neomycin and polymyxin B for?
Neomycin, polymyxin B, and dexamethasone combination is used to treat eye infections and inflammation, including conjunctivitis and chronic anterior uveitis. It also prevents damage caused by chemicals, radiation, or foreign objects entering the eye.
What is polymyxin B used for?
Polymyxin B and trimethoprim combination is used to treat eye infections, including acute bacterial conjunctivitis and blepharoconjunctivitis.
How is the hygromycin B phosphotransferase (HPH) coding sequence determined?
The nucleotide sequence of a 1467 bp fragment of Streptomyces hygroscopicus DNA containing the gene (hyg) encoding a hygromycin B phosphotransferase (HPH) has been determined. The N-ter-minal atnino acid sequence of HPH determined by automated Edman degradation has allowed the coding sequence of the hyg gene to be identified.
Is there a hygromycin B resistance gene in Escherichia coli?
Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae The plasmid-borne gene hph coding for hygromycin B phosphotransferase (HPH) in Escherichia coli has been identified and its nucleotide sequence determined.
What is hygromycin B made from?
Hygromycin B is produced by Streptomyces hygroscopicus, a bacterium isolated in 1953 from a soil sample. Resistance genes were discovered in the early 1980s. Hygromycin is active against both prokaryotic and eukaryotic cells. It acts by inhibiting polypeptide synthesis.
Is hygromycin B a good antibiotic for gene transfer?
Since the discovery of hygromycin-resistance genes, hygromycin B has become a standard selection antibiotic in gene transfer experiments in many prokaryotic and eukaryotic cells.
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