Abstract
A novel form of "enzyme therapy" was achieved by utilizing protoplasts of Bacillus subtilis. Photoreactivating enzyme of Escherichia coli was successfully inserted into the protoplasts of B. subtilis treated with polyethylene glycol. This enzyme was used to photoreactivate ultraviolet-damaged bacteriophage deoxyribonucleic acid (DNA). Furthermore, in polyethylene glycol-treated protoplasts, ultraviolet-irradiated transfecting bacteriophage DNA was shown to be a functional substrate for the host DNA excision repair system. Previous results (R. E. Yasbin, J. D. Fernwalt, and P. I. Fields, J. Bacteriol. 137:391-396, 1979) showed that ultraviolet-irradiated bacteriophage DNA could not be repaired via the excision repair system of competent cells. Therefore, the processing of bacteriophage DNA by protoplasts and by competent cells must be different. This sensitive protoplast assay can be used to identify and to isolate various types of DNA repair enzymes.
Full text
PDF




Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Campbell L. A., Yasbin R. E. Deoxyribonucleic acid repair capacities of Neisseria gonorrhoeae: absence of photoreactivation. J Bacteriol. 1979 Dec;140(3):1109–1111. doi: 10.1128/jb.140.3.1109-1111.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
- Hadden C. T. Postirradiation recovery dependent on the uvr-1 locus in Bacillus subtilis. J Bacteriol. 1976 Oct;128(1):317–324. doi: 10.1128/jb.128.1.317-324.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jagger J., Stafford R. S., Snow J. M. Thymine-dimer and action-spectrum evidence for indirect photoreactivation in Escherichia coli. Photochem Photobiol. 1969 Dec;10(6):383–395. doi: 10.1111/j.1751-1097.1969.tb05703.x. [DOI] [PubMed] [Google Scholar]
- Lovett P. S., Keggins K. M. Bacillus subtilis as a host for molecular cloning. Methods Enzymol. 1979;68:342–357. doi: 10.1016/0076-6879(79)68025-4. [DOI] [PubMed] [Google Scholar]
- Nester E W, Schafer M, Lederberg J. Gene Linkage in DNA Transfer: A Cluster of Genes Concerned with Aromatic Biosynthesis in Bacillus Subtilis. Genetics. 1963 Apr;48(4):529–551. doi: 10.1093/genetics/48.4.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rupert C. S. Enzymatic photoreactivation: overview. Basic Life Sci. 1975;5A:73–87. doi: 10.1007/978-1-4684-2895-7_11. [DOI] [PubMed] [Google Scholar]
- Snapka R. M., Sutherland B. M. Escherichia coli photoreactivating enzyme: purification and properties. Biochemistry. 1980 Sep 2;19(18):4201–4208. doi: 10.1021/bi00559a010. [DOI] [PubMed] [Google Scholar]
- Sutherland B. M., Hausrath S. G. Insertion of E. coli photoreactivating enzyme into V79 hamster cells. Nature. 1980 Jul 31;286(5772):510–511. doi: 10.1038/286510a0. [DOI] [PubMed] [Google Scholar]
- White T. B., Doyle R. J., Streips U. N. Transformation of a Bacillus subtilis L-form with bacteriophage deoxyribonucleic acid. J Bacteriol. 1981 Feb;145(2):878–883. doi: 10.1128/jb.145.2.878-883.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yasbin R. E. DNA repair in Bacillus subtilis. II. Activation of the inducible system in competent bacteria. Mol Gen Genet. 1977 Jun 8;153(2):219–225. [PubMed] [Google Scholar]
- Yasbin R. E., Fernwalt J. D., Fields P. I. DNA repair in Bacillus subtilis: excision repair capacity of competent cells. J Bacteriol. 1979 Jan;137(1):391–396. doi: 10.1128/jb.137.1.391-396.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yasbin R. E., Fields P. I., Andersen B. J. Properties of Bacillus subtilis 168 derivatives freed of their natural prophages. Gene. 1980 Dec;12(1-2):155–159. doi: 10.1016/0378-1119(80)90026-8. [DOI] [PubMed] [Google Scholar]
- Yasbin R. E., Miehl R. Deoxyribonucleic Acid Repair in Bacillus subtilis: Development of Competent Cells into a Tester for Carcinogens. Appl Environ Microbiol. 1980 Apr;39(4):854–858. doi: 10.1128/aem.39.4.854-858.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yasbin R. E., Wilson G. A., Young F. E. Transformation and transfection in lysogenic strains of Bacillus subtilis 168. J Bacteriol. 1973 Feb;113(2):540–548. doi: 10.1128/jb.113.2.540-548.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yasbin R. E., Wilson G. A., Young F. E. Transformation and transfection in lysogenic strains of Bacillus subtilis: evidence for selective induction of prophage in competent cells. J Bacteriol. 1975 Jan;121(1):296–304. doi: 10.1128/jb.121.1.296-304.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
