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Incorporation of cloned DNA by Bacillus subtilis

Technical Report ·
OSTI ID:6471381
Chimeric plasmids, consisting of Bacillus DNA coupled to Escherichia coli DNA, were used to study genetic exchange in the soil bacterium, Basillus subtilis. These studies revealed that plasmids containing thyP3, a gene which shares nucleotide sequence homology with the B. subtilis chromosome, transform B. subtilis thymine auxotrophs to prototrophy at a high frequency whether the plasmid DNA is in circular or linear form. Transformation of the same strain with plasmids containing the gene thy..beta..22 occurred at a much lower frequency, required circular DNA molecules, and required the presence of DNA sequences homologous to the B. subtilis chromosome on the plasmid. Hybridization analysis of chromosomal DNA from clones transformed with thy..beta..22 demonstrated that the entire chimeric plasmid was inserted in the chromosome by a Campbell style integration event. This type of additive recombination had not been previously observed during transformation of B. subtilis. To facilitate this work the two site specific endonucleases from Bacillus globigii were characterized by genetic and biochemical procedures. These studies led to improved purification techniques for the two enzymes.
Research Organization:
Rochester Univ., NY (USA). Dept. of Radiation Biology
DOE Contract Number:
EY-76-C-02-3490
OSTI ID:
6471381
Report Number(s):
UR-3490/LCP-14
Country of Publication:
United States
Language:
English