Molecular cloning and characterization of comC, a late competence gene of Bacillus subtilis

S. Mohan, J. Aghion, N. Guillen, David Dubnau

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

comC is a Bacillus subtilis gene required for the development of genetic competence. We have cloned a fragment from the B. subtilis chromosome that carries comC and contains all the information required to complement a Tn917lac insertion in comC. Genetic tests further localized comC to a 2.0-kilobase HindIII fragment. Northern (RNA) blotting experiments revealed that an 800-base-pair comC-specific transcript appeared at the time of transition from exponential to stationary phase during growth through the competence regimen. The DNA sequence of the comC region revealed two open reading frames (ORFs), transcribed in the same direction. The upstream ORF encoded a protein with apparent sequence similarity to the folC gene of Escherichia coli. Insertion of a chloramphenicol resistance determinant into this ORF and integration of the disrupted construct into the bacterial chromosome by replacement did not result in competence deficiency. The downstream ORF, which contained the Tn917lac insertion that resulted in a lack of competence, is therefore the comC gene. The predicted protein product of comC consisted of 248 amino acid residues and was quite hydrophobic. The comC gene product was not required for the expression of any other com genes tested, and this fact, together with the marked hydrophobicity of ComC, suggests that it may be a component of the DNA-processing apparatus of competent cells.

Original languageEnglish (US)
Pages (from-to)6043-6051
Number of pages9
JournalJournal of bacteriology
Volume171
Issue number11
DOIs
StatePublished - Jan 1 1989

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Molecular Cloning
Bacillus subtilis
Mental Competency
Open Reading Frames
Genes
Bacterial Chromosomes
Chloramphenicol Resistance
Hydrophobic and Hydrophilic Interactions
Base Pairing
Northern Blotting
Proteins
Chromosomes
RNA
Escherichia coli
Amino Acids
DNA
Growth

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Mohan, S. ; Aghion, J. ; Guillen, N. ; Dubnau, David. / Molecular cloning and characterization of comC, a late competence gene of Bacillus subtilis. In: Journal of bacteriology. 1989 ; Vol. 171, No. 11. pp. 6043-6051.
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Molecular cloning and characterization of comC, a late competence gene of Bacillus subtilis. / Mohan, S.; Aghion, J.; Guillen, N.; Dubnau, David.

In: Journal of bacteriology, Vol. 171, No. 11, 01.01.1989, p. 6043-6051.

Research output: Contribution to journalArticle

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AB - comC is a Bacillus subtilis gene required for the development of genetic competence. We have cloned a fragment from the B. subtilis chromosome that carries comC and contains all the information required to complement a Tn917lac insertion in comC. Genetic tests further localized comC to a 2.0-kilobase HindIII fragment. Northern (RNA) blotting experiments revealed that an 800-base-pair comC-specific transcript appeared at the time of transition from exponential to stationary phase during growth through the competence regimen. The DNA sequence of the comC region revealed two open reading frames (ORFs), transcribed in the same direction. The upstream ORF encoded a protein with apparent sequence similarity to the folC gene of Escherichia coli. Insertion of a chloramphenicol resistance determinant into this ORF and integration of the disrupted construct into the bacterial chromosome by replacement did not result in competence deficiency. The downstream ORF, which contained the Tn917lac insertion that resulted in a lack of competence, is therefore the comC gene. The predicted protein product of comC consisted of 248 amino acid residues and was quite hydrophobic. The comC gene product was not required for the expression of any other com genes tested, and this fact, together with the marked hydrophobicity of ComC, suggests that it may be a component of the DNA-processing apparatus of competent cells.

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