Chromosomal-DNA Amplification in Bacillus Subtilis

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1985 Aug 1

PMID 2991188

Citations 14

Authors

C R Wilson

A E Morgan

Affiliations

Soon will be listed here.

Abstract

Tetracycline-resistant (Tetr) mutants RAD1, RAD2, RAD6, and RAD7 were isolated from Bacillus subtilis BC92 after protoplasting, polyethylene glycol treatment, and regeneration on a medium containing tetracycline. The Tetr phenotype in RAD1, RAD2, and RAD6 was very stable with less than 5% loss of resistance after 30 generations of growth in the absence of selection. Of the four isolates, three contained amplified chromosomal DNA closely associated with the Tetr phenotype. The intensity of restriction fragments present in HindIII and EcoRI digests of chromosomal DNA from RAD1, RAD6, and RAD7 indicated the presence of tandemly duplicated DNA. Disparity in the size and number of amplified fragments suggested that the tandemly duplicated DNA is different in all three isolates. The sizes of the duplicated DNA present in RAD1, RAD6, and RAD7 were estimated to be 10, 19, and 20 kilobases, respectively. No amplified DNA was detected in RAD2. Results of transductional-mapping studies with PBS1 showed that the tetracycline resistance (tet) loci of RAD1, RAD2, and RAD6 all mapped near the origin of chromosomal replication and close to the guaA locus. Amplified DNA characteristic of RAD1 and RAD6 was cotransduced with the tet locus. Cotransfer of amplified DNA with the guaA locus or other nearby loci in the absence of tet was not observed. In every case, loss of Tetr was accompanied by loss of amplified DNA. A possible explanation for the occurrence of the amplified DNA is presented.

Citing Articles

On the mechanism of gene amplification induced under stress in Escherichia coli.

Slack A, Thornton P, Magner D, Rosenberg S, Hastings P PLoS Genet. 2006; 2(4):e48.

PMID: 16604155 PMC: 1428787. DOI: 10.1371/journal.pgen.0020048.

Tetracycline induces stabilization of mRNA in Bacillus subtilis.

Wei Y, Bechhofer D J Bacteriol. 2002; 184(4):889-94.

PMID: 11807047 PMC: 134818. DOI: 10.1128/jb.184.4.889-894.2002.

tetA(L) mutants of a tetracycline-sensitive strain of Bacillus subtilis with the polynucleotide phosphorylase gene deleted.

Bechhofer D, Stasinopoulos S J Bacteriol. 1998; 180(13):3470-3.

PMID: 9642204 PMC: 107306. DOI: 10.1128/JB.180.13.3470-3473.1998.

Tetracycline/H+ antiport and Na+/H+ antiport catalyzed by the Bacillus subtilis TetA(L) transporter expressed in Escherichia coli.

Guffanti A, Krulwich T J Bacteriol. 1995; 177(15):4557-61.

PMID: 7635843 PMC: 177215. DOI: 10.1128/jb.177.15.4557-4561.1995.

Transformation of the cyanobacterium Anacystis nidulans 6301 with the Escherichia coli plasmid pBR322.

Daniell H, Sarojini G, McFadden B Proc Natl Acad Sci U S A. 1986; 83(8):2546-50.

PMID: 3085098 PMC: 323335. DOI: 10.1073/pnas.83.8.2546.

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