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PMID: 6260747 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation of Bacillus subtilis genes from a charon 4A library.

Journal of bacteriology ·Vol. 146 ·No. 1 ·1981-04-00 ·Pages 430-2

Ferrari E, Henner DJ, Hoch JA

Abstract

A library of Bacillus subtilis chromosomal deoxyribonucleic acid (DNA) was constructed, using lambda charon 4A as a cloning vector. Partially cleaved Bacillus subtilis DNA was prepared by partial methylation with EcoRI methylase, followed by complete EcoRI endonuclease digestion. More than 95% of the phage particles carried B. subtilis DNA inserts. When this library was screened for transforming activity, using competent cells, 70% of the genetic markers tested were found in a sample of 1,710 plaques. Cloned genetic loci were found to be about 100-fold more efficient in transforming activity than chromosomal DNA. Intact phage particles containing the pheA locus were found to be able to transform competent recipients with approximately the same efficiency as phage DNA. Transformation by intact particles was insensitive to deoxyribonuclease.

MeSH Terms

Bacillus subtilis/genetics Bacteriophages Chromosome Mapping Cloning, Molecular DNA Restriction Enzymes DNA, Bacterial/genetics Genetic Markers Methyltransferases Site-Specific DNA-Methyltransferase (Adenine-Specific) Transformation, Bacterial

Chemicals

DNA, Bacterial Genetic Markers DNA modification methylase EcoRI Methyltransferases Site-Specific DNA-Methyltransferase (Adenine-Specific) DNA Restriction Enzymes

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Ferrari E

Henner D J

Hoch J A

References (10)

10 references, click to expand

REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS.
J Bacteriol. 1961 May;81(5):741-6 PMID: 16561900
Restriction enzyme analysis of Bacillus subtilis ribosomal ribonucleic acid genes.
J Bacteriol. 1979 Oct;140(1):99-105 PMID: 115842
Construction of a colony bank of E. coli containing hybrid plasmids representative of the Bacillus subtilis 168 genome. Expression of functions harbored by the recombinant plasmids in B. subtilis.
Mol Gen Genet. 1979 Oct 3;176(2):239-45 PMID: 119126
Packaging recombinant DNA molecules into bacteriophage particles in vitro.
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3259-63 PMID: 333431
A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome.
Cell. 1976 Sep;9(1):91-9 PMID: 788919
The isolation of structural genes from libraries of eucaryotic DNA.
Cell. 1978 Oct;15(2):687-701 PMID: 719759
Cloning of randomly sheared DNA fragments from a phi 105 lysogen of Bacillus subtilis identification of prophage-containing clones.
Gene. 1980 Feb;8(3):267-78 PMID: 6766891
Charon phages: safer derivatives of bacteriophage lambda for DNA cloning.
Science. 1977 Apr 8;196(4286):161-9 PMID: 847462
Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease.
J Mol Biol. 1975 Jan 25;91(3):315-28 PMID: 1102702
Screening lambdagt recombinant clones by hybridization to single plaques in situ.
Science. 1977 Apr 8;196(4286):180-2 PMID: 322279

Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.146.1.430-432.1981

Published

1981-04-00

Pages

430-2

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC217104

Subset

Grants

NIGMS NIH HHS · GM07576 · United States

NIGMS NIH HHS · GM19416 · United States

NIGMS NIH HHS · GM25891 · United States

External Links

PubMed source DOI: 10.1128/jb.146.1.430-432.1981 PMC Full Text View journal details

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Isolation of Bacillus subtilis genes from a charon 4A library.

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