Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

M S Sørensen; M Duch; K Paludan; Poul Jørgensen; F S Pedersen

Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

M S Sørensen, M Duch, K Paludan, Poul Jørgensen, F S Pedersen

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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Abstract

Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.

Original language	English
Journal	Gene
Volume	112
Issue	2
Pages (from-to)	257-60
Number of pages	3
ISSN	0378-1119
Publication status	Published - 1992

Keywords

Animals
Cell Extracts
Cell Line
Genetic Markers
Kanamycin Kinase
Phosphotransferases
Phosphotransferases (Alcohol Group Acceptor)
Substrate Specificity

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@article{5fa84150ab1811dd889c000ea68e967b,

title = "Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay",

abstract = "Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.",

keywords = "Animals, Cell Extracts, Cell Line, Genetic Markers, Kanamycin Kinase, Phosphotransferases, Phosphotransferases (Alcohol Group Acceptor), Substrate Specificity",

author = "S{\o}rensen, {M S} and M Duch and K Paludan and Poul J{\o}rgensen and Pedersen, {F S}",

year = "1992",

language = "English",

volume = "112",

pages = "257--60",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier BV",

number = "2",

}

TY - JOUR

T1 - Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

AU - Sørensen, M S

AU - Duch, M

AU - Paludan, K

AU - Jørgensen, Poul

AU - Pedersen, F S

PY - 1992

Y1 - 1992

N2 - Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.

AB - Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.

KW - Animals

KW - Cell Extracts

KW - Cell Line

KW - Genetic Markers

KW - Kanamycin Kinase

KW - Phosphotransferases

KW - Phosphotransferases (Alcohol Group Acceptor)

KW - Substrate Specificity

M3 - Journal article

C2 - 1313387

SN - 0378-1119

VL - 112

SP - 257

EP - 260

JO - Gene

JF - Gene

IS - 2

ER -

Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

Abstract

Keywords

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