The illegitimate recombination between Staphylococcus aureus plasmids pE194 (or pGG20, the hybrid between pE194 and Escherichia coli plasmid pBR322) and pBD17 (plasmid pUB110 without HpaII C-fragment) was studied in Bacillus subtilis. Cointegrates were generated with the frequency of 1-3 X 10(-8). A
Illegitimate recombination in Bacillus subtilis: nucleotide sequences at recombinant DNA junctions
β Scribed by Bashkirov, Vladimir I. ;Khasanov, Fuat K. ;Prozorov, Alexander A.
- Book ID
- 104723418
- Publisher
- Springer
- Year
- 1987
- Tongue
- English
- Weight
- 375 KB
- Volume
- 210
- Category
- Article
- ISSN
- 0026-8925
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β¦ Synopsis
The illegitimate integration of plasmid pGG20 (the hybrid between Staphylococcus aureus plasmid pE194 and Escherichia coli plasmid pBR322) into the Bacillus subtilis chromosome was studied. It was found that nucleotide sequences of both parental plasmids could be involved in this process. The recombinant DNA junctions between plasmid pGG20 and the chromosome were cloned and their nucleotide sequences were determined. The site of recombination located on the pBR322 moiety carried a short region (8 bp) homologous with the site on the chromosome. The nucleotide sequences of the pE194 recombination sites did not share homology with chromosomal sequences involved in the integration process. Two different pathways of illegitimate recombination in B. subtilis are suggested.
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Directly repeated pBR322 sequences 3.7-3.8 kb long recombine with a frequency of about 10% per generation when carried on plasmids related to pC194 and 0.01% per generation when carried on the Bacillus subtilis chromosome. Recombination is therefore 1,000 times more efficient in plasmids than in the
DNA of Bacillus subtilis strain UVSS 19--8M, of high ultraviolet sensitivity, was isolated after cultivating in medium containing bromouracil. Isopycnic banding in CsC1 shows an unusual pattern with four bands, including an extra one halfway between those for hybrid and for DNA of this band, amounti