Cloning of the Bacillus subtilis recE/sup +/ gene and functional expression of recE/sup +/ in B. subtilis

Journal Article · Fri Jan 01 04:00:00 EST 1988 · J. Bacteriol.; (United States)

OSTI ID:5272205

Marrero, R; Yasbin, R E

By use of the Bacillus subtilis bacteriophage cloning vehicle Phi 105J23, B. subtilis chromosomal MboI fragments have been cloned that alleviate the pleiotropic effects of the recE4 mutation. The recombinant bacteriophages Phi 105Rec Phi1 (3.85-kilobase insert) and Phi 105Rec Phi4 (3.3-kilobase insert) both conferred on the recE4 strain YB1015 resistance to ethylmethane sulfonate, methylmethane sulfonate, mitomycin C, and UV irradiation comparable with the resistance observed in recE/sup +/ strains. While strain YB1015 (recE4) and its derivatives lysogenized with bacteriophage Phi105J23 were not transformed to prototrophy by B. subtilis chromosomal DNA, strain YB1015 lysogenized with either Phi 105Rec Phi 1 or Phi 105RecPhi 4 was susceptible to transformation with homologous B. subtilis chromosomal DNA. The heteroimmune prophages Phi 105 and SPO2 were essentially uninducible in strain YB1015. Significantly, both recombinant prophages Phi 105RecPhi 1 and Phi 105Rec Phi 4 were fully inducible and allowed the spontaneous and mitomycin C-dependent induction of a coresident SPO2 prophage in a recE4 host. The presence of the recombinant prophages also restored the ability of din genes to be induced in strains carrying the recE4 mutation. Finally, both recombinant bacteriophages elaborated a mitomycin C-inducible, 45-kilodalton protein that was immunoreactive with Escherichia coli recA/sup +/ gene product antibodies. Collectively, these data demonstrate that the recE/sup +/ gene has been cloned and that this gene elaborates the 45-kilodalton protein that is involved in SOB induction and homologous recombination.

• Research Organization: Univ. of Rochester School of Medicine and Dentistry, NY
• DOE Contract Number: AC02-81ER60017
• OSTI ID: 5272205
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 170:1; ISSN JOBAA
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ANTIMITOTIC DRUGS
ANTINEOPLASTIC DRUGS
BACILLUS
BACILLUS SUBTILIS
BACTERIA
BACTERIOPHAGES
CLONING
DNA
DNA-CLONING
DRUGS
ELECTROMAGNETIC RADIATION
EMS
ESTERS
GENES
GENETIC MAPPING
MAPPING
METHYL METHANESULFONATE
MICROORGANISMS
MITOMYCIN
MOLECULAR BIOLOGY
MUTAGENESIS
MUTAGENS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PARASITES
PROTEINS
RADIATIONS
RECOMBINANT DNA
SULFONIC ACID ESTERS
ULTRAVIOLET RADIATION
VIRUSES