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PMID: 96091 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Recombination between compatible plasmids containing homologous segments requires the Bacillus subtilis recE gene product.

Journal of bacteriology ·Vol. 134 ·No. 2 ·1978-05-00 ·Pages 514-20

Keggins KM, Duvall EJ, Lovett PS

Abstract

Plasmid pSL103 was previously constructed by cloning a Trp fragment (approximately 2.3 X 10(6) daltons) from restriction endonuclease EcoRI-digested chromosome DNA of Bacillus pumilus using the neomycin-resistance plasmid pUB110 (approximately 2.8 X 10(6) daltons) as vector and B. subtilis as transformation recipient. In the present study the EcoRI Trp fragment from pSL103 was transferred in vitro to EcoRI fragments of the Bacillus plasmid pPL576 to determine the ability of the plasmid fragments to replicate in B. subtilis. Endonuclease EcoRI digestion of pPL576 (approximately 28 X 10(6) daltons) generated three fragments having molecular weights of about 13 X 13(6) (the A fragment), 9.5 X 10(6) (B fragment, and 6.5 X 10(6) (C fragment). Trp derivatives of pPL576 fragments capable of autonomous replication in B. subtilis contained the B fragment (e.g., pSL107) or both the B and C fragments (e.g., pSL108). Accordingly, the B fragment of pPL576 contains information essential for autonomous replication. pSL107 and pSL108 are compatible with pUB110. Constructed derivatives of the compatible plasmids pPL576 and pUB110, harboring genetically distinguishable EcoRI-generated Trp fragments cloned from the DNA of a B. pumilus strain, exhibited relatively high frequency recombination for a trpC marker when the plasmid pairs were present in a recombination-proficient strain of B. subtilis. No recombination was detected when the host carried the chromosome mutation recE4. Therefore, the recE4 mutation suppresses recombination between compatible plasmids that contain homologous segments.

MeSH Terms
Bacillus subtilis/genetics Chromosomes, Bacterial DNA Replication DNA, Bacterial/metabolism DNA, Recombinant/metabolism Genes Mutation Plasmids Recombination, Genetic Transformation, Bacterial
Chemicals
DNA, Bacterial DNA, Recombinant
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Keggins K M
Duvall E J
Lovett P S
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25 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1978-05-00
Pages
514-20
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC222281
Subset
IM
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