A genetic system for analysis of staphylococcal nuclease

David Shortle

doi:10.1016/0378-1119(83)90102-6

A genetic system for analysis of staphylococcal nuclease

David Shortle

Research output: Contribution to journal › Article › peer-review

159 Scopus citations

Abstract

The gene encoding staphylococcal nuclease (strain Foggi) has been isolated, and its complete nucleotide sequence determined. When inserted into a derivative of plasmid pBR322, the nuc gene is expressed in Escherichia coli at a low level, and nuclease activity in individual colonies is readily assayed by a replica-plating chromogenic test. A set of plasmids with BamHI-linker "bracketed" deletions spanning the nuc gene can be used to map mutations genetically to defined segments of the gene. In combination with present methods for efficient in vitro mutagenesis, this plasmid-based genetic system can be applied to the detailed genetic analysis of this small, biochemically and biophysically well-characterized enzyme.

Original language	English (US)
Pages (from-to)	181-189
Number of pages	9
Journal	Gene
Volume	22
Issue number	2-3
DOIs	https://doi.org/10.1016/0378-1119(83)90102-6
State	Published - 1983
Externally published	Yes

Keywords

Recombinant DNA
deletion mapping
enzyme genetics
linker mutagenesis
micrococcal nuclease
restriction map
signal sequence
ucleotide sequence

ASJC Scopus subject areas

Genetics

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10.1016/0378-1119(83)90102-6

Cite this

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title = "A genetic system for analysis of staphylococcal nuclease",

abstract = "The gene encoding staphylococcal nuclease (strain Foggi) has been isolated, and its complete nucleotide sequence determined. When inserted into a derivative of plasmid pBR322, the nuc gene is expressed in Escherichia coli at a low level, and nuclease activity in individual colonies is readily assayed by a replica-plating chromogenic test. A set of plasmids with BamHI-linker {"}bracketed{"} deletions spanning the nuc gene can be used to map mutations genetically to defined segments of the gene. In combination with present methods for efficient in vitro mutagenesis, this plasmid-based genetic system can be applied to the detailed genetic analysis of this small, biochemically and biophysically well-characterized enzyme.",

keywords = "Recombinant DNA, deletion mapping, enzyme genetics, linker mutagenesis, micrococcal nuclease, restriction map, signal sequence, ucleotide sequence",

author = "David Shortle",

note = "Funding Information: I would like to thank Andrew Ellington for helpful discussions, Dan Davison for his computer analysis of the DNA sequence. Sandi Burns for typing the manuscript and Patrick Hearing, Tom Shenk, and Jim Broach for helpful comments on the manuscript. Financial support for this research was from NIH Grant GM 306-4901.",

year = "1983",

doi = "10.1016/0378-1119(83)90102-6",

language = "English (US)",

volume = "22",

pages = "181--189",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "2-3",

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T1 - A genetic system for analysis of staphylococcal nuclease

AU - Shortle, David

N1 - Funding Information: I would like to thank Andrew Ellington for helpful discussions, Dan Davison for his computer analysis of the DNA sequence. Sandi Burns for typing the manuscript and Patrick Hearing, Tom Shenk, and Jim Broach for helpful comments on the manuscript. Financial support for this research was from NIH Grant GM 306-4901.

PY - 1983

Y1 - 1983

N2 - The gene encoding staphylococcal nuclease (strain Foggi) has been isolated, and its complete nucleotide sequence determined. When inserted into a derivative of plasmid pBR322, the nuc gene is expressed in Escherichia coli at a low level, and nuclease activity in individual colonies is readily assayed by a replica-plating chromogenic test. A set of plasmids with BamHI-linker "bracketed" deletions spanning the nuc gene can be used to map mutations genetically to defined segments of the gene. In combination with present methods for efficient in vitro mutagenesis, this plasmid-based genetic system can be applied to the detailed genetic analysis of this small, biochemically and biophysically well-characterized enzyme.

AB - The gene encoding staphylococcal nuclease (strain Foggi) has been isolated, and its complete nucleotide sequence determined. When inserted into a derivative of plasmid pBR322, the nuc gene is expressed in Escherichia coli at a low level, and nuclease activity in individual colonies is readily assayed by a replica-plating chromogenic test. A set of plasmids with BamHI-linker "bracketed" deletions spanning the nuc gene can be used to map mutations genetically to defined segments of the gene. In combination with present methods for efficient in vitro mutagenesis, this plasmid-based genetic system can be applied to the detailed genetic analysis of this small, biochemically and biophysically well-characterized enzyme.

KW - Recombinant DNA

KW - deletion mapping

KW - enzyme genetics

KW - linker mutagenesis

KW - micrococcal nuclease

KW - restriction map

KW - signal sequence

KW - ucleotide sequence

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DO - 10.1016/0378-1119(83)90102-6

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AN - SCOPUS:0020583027

SN - 0378-1119

VL - 22

SP - 181

EP - 189

JO - Gene

JF - Gene

IS - 2-3

ER -

A genetic system for analysis of staphylococcal nuclease

Abstract

Keywords

ASJC Scopus subject areas

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