Abstract
Both fi+ and fi− type R factors function as intact replicative units when segregated into Escherichia coli minicells. Hybridization studies demonstrate that at least 95% of the deoxyribonucleic acid (DNA) in R+ minicells is episomal in origin. About half of the DNA can be extracted in a closed circular form and about 75% of the DNA is membrane associated. DNA, ribonucleic acid, and protein synthesis proceeds in R+ minicells in contrast to R− minicell controls. The system offers a unique opportunity to study a relatively small replicative unit in a native cell environment and a simple means of isolating large quantities of episomal DNA.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Adesnik M., Levinthal C. Synthesis and maturation of ribosomal RNA in Escherichia coli. J Mol Biol. 1969 Dec 14;46(2):281–303. doi: 10.1016/0022-2836(69)90422-7. [DOI] [PubMed] [Google Scholar]
- Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BERNS K. I., THOMAS C. A., Jr ISOLATION OF HIGH MOLECULAR WEIGHT DNA FROM HEMOPHILUS INFLUENZAE. J Mol Biol. 1965 Mar;11:476–490. doi: 10.1016/s0022-2836(65)80004-3. [DOI] [PubMed] [Google Scholar]
- BOLTON E. T., McCARTHY B. J. A general method for the isolation of RNA complementary to DNA. Proc Natl Acad Sci U S A. 1962 Aug;48:1390–1397. doi: 10.1073/pnas.48.8.1390. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brenner D. J., Fanning G. R., Johnson K. E., Citarella R. V., Falkow S. Polynucleotide sequence relationships among members of Enterobacteriaceae. J Bacteriol. 1969 May;98(2):637–650. doi: 10.1128/jb.98.2.637-650.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CURTIS S. R., 3rd CHROMOSOMAL ABERRATIONS ASSOCIATED WITH MUTATIONS TO BACTERIOPHAGE RESISTANCE IN ESCHERICHIA COLI. J Bacteriol. 1965 Jan;89:28–40. doi: 10.1128/jb.89.1.28-40.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen A., Fisher W. D., Curtiss R., 3rd, Adler H. I. The properties of DNA transferred to minicells during conjugation. Cold Spring Harb Symp Quant Biol. 1968;33:635–641. doi: 10.1101/sqb.1968.033.01.071. [DOI] [PubMed] [Google Scholar]
- Cohen S. N., Miller C. A. Multiple molecular species of circular R-factor DNA isolated from Escherichia coli. Nature. 1969 Dec 27;224(5226):1273–1277. doi: 10.1038/2241273a0. [DOI] [PubMed] [Google Scholar]
- Falkow S., Citarella R. V., Wohlhieter J. A. The molecular nature of R-factors. J Mol Biol. 1966 May;17(1):102–116. doi: 10.1016/s0022-2836(66)80097-9. [DOI] [PubMed] [Google Scholar]
- Freifelder D. Studies on Escherichia coli sex factors. IV. Molecular weights of the DNA of several F' elements. J Mol Biol. 1968 Jul 14;35(1):95–102. doi: 10.1016/s0022-2836(68)80039-7. [DOI] [PubMed] [Google Scholar]
- Hartmann G., Honikel K. O., Knüsel F., Nüesch J. The specific inhibition of the DNA-directed RNA synthesis by rifamycin. Biochim Biophys Acta. 1967;145(3):843–844. doi: 10.1016/0005-2787(67)90147-5. [DOI] [PubMed] [Google Scholar]
- Inselburg J. R factor deoxyribonucleic acid in chromosomeless progeny of Escherichia coli. J Bacteriol. 1971 Feb;105(2):620–628. doi: 10.1128/jb.105.2.620-628.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inselburg J. Segregation into and replication of plasmid deoxyribonucleic acid in chromosomeless segregants of Escherichia coli. J Bacteriol. 1970 Jun;102(3):642–647. doi: 10.1128/jb.102.3.642-647.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kass L. R., Yarmolinsky M. B. Segregation of functional sex factor into minicells. Proc Natl Acad Sci U S A. 1970 Jul;66(3):815–822. doi: 10.1073/pnas.66.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knüsel F., Schiess B. Dominance study with rifampicin-resistant mutants of E. coli K 12. Mol Gen Genet. 1970;108(4):331–337. doi: 10.1007/BF00267770. [DOI] [PubMed] [Google Scholar]
- LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
- Levy S. B., Norman P. Segregation of transferable R factors into Escherichia coli minicells. Nature. 1970 Aug 8;227(5258):606–607. doi: 10.1038/227606a0. [DOI] [PubMed] [Google Scholar]
- Levy S. B. Resistance of minicells to penicillin lysis: a method of obtaining large quantities of purified minicells. J Bacteriol. 1970 Sep;103(3):836–839. doi: 10.1128/jb.103.3.836-839.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levy S. B. The problems of drug-resistant pathogenic bacteria. Studies on R factors segregated into E. coli minicells. Ann N Y Acad Sci. 1971 Jun 11;182:217–225. doi: 10.1111/j.1749-6632.1971.tb30658.x. [DOI] [PubMed] [Google Scholar]
- Meynell E., Datta N. Mutant drug resistant factors of high transmissibility. Nature. 1967 May 27;214(5091):885–887. doi: 10.1038/214885a0. [DOI] [PubMed] [Google Scholar]
- Meynell E., Meynell G. G., Datta N. Phylogenetic relationships of drug-resistance factors and other transmissible bacterial plasmids. Bacteriol Rev. 1968 Mar;32(1):55–83. doi: 10.1128/br.32.1.55-83.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nisioka T., Mitani M., Clowes R. Composite circular forms of R factor deoxyribonucleic acid molecules. J Bacteriol. 1969 Jan;97(1):376–385. doi: 10.1128/jb.97.1.376-385.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rownd R., Nakaya R., Nakamura A. Molecular nature of the drug-resistance factors of the Enterobacteriaceae. J Mol Biol. 1966 Jun;17(2):376–393. doi: 10.1016/s0022-2836(66)80149-3. [DOI] [PubMed] [Google Scholar]
- Rownd R. Replication of a bacterial episome under relaxed control. J Mol Biol. 1969 Sep 28;44(3):387–402. doi: 10.1016/0022-2836(69)90368-4. [DOI] [PubMed] [Google Scholar]
- Silver R. P., Falkow S. Specific labeling and physical characterization of R-factor deoxyribonucleic acid in Escherichia coli. J Bacteriol. 1970 Oct;104(1):331–339. doi: 10.1128/jb.104.1.331-339.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
- WATANABE T. Infective heredity of multiple drug resistance in bacteria. Bacteriol Rev. 1963 Mar;27:87–115. doi: 10.1128/br.27.1.87-115.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wehrli W., Knüsel F., Schmid K., Staehelin M. Interaction of rifamycin with bacterial RNA polymerase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):667–673. doi: 10.1073/pnas.61.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young R. W., Fulhorst H. W. Recovery of S35 radioactivity from protein-bearing polyacrylamide gel. Anal Biochem. 1965 May;11(2):389–391. doi: 10.1016/0003-2697(65)90030-8. [DOI] [PubMed] [Google Scholar]