Abstract
Transposon Tn5 conferred streptomycin resistance on different strains of Rhizobium meliloti, Rhizobium leguminosarum, and Rhizobium trifolii but not on Escherichia coli. A gene (str) specifying this phenotype has been identified and localized on the physical and genetic map of Tn5. It is transcribed from the promoter of neo, the gene that encodes neomycin phosphotransferase. The str gene is downstream from neo in a single transcriptional unit, as revealed by molecular cloning of different segments of Tn5 and by cloning of the neo-str region of Tn5 downstream from a lac promoter. Fusion of the SalI-generated rightward segment of Tn5 (devoid of neo) to a part of a tetracycline resistance gene, tet, in a plasmid or downstream from a lac promoter in a plasmid resulted in significant levels of streptomycin resistance in an R. meliloti host, suggesting that the str gene product can function independent of neomycin phosphotransferase. A natural isolate of R. meliloti that does not express Tn5-associated streptomycin resistance has been identified. We have used the str of Tn5 as a genetic marker in Rhizobium spp.
Full text
PDF









Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Auerswald E. A., Ludwig G., Schaller H. Structural analysis of Tn5. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):107–113. doi: 10.1101/sqb.1981.045.01.019. [DOI] [PubMed] [Google Scholar]
- Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
- Berg D. E. Control of gene expression by a mobile recombinational switch. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4880–4884. doi: 10.1073/pnas.77.8.4880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berg D. E., Davies J., Allet B., Rochaix J. D. Transposition of R factor genes to bacteriophage lambda. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3628–3632. doi: 10.1073/pnas.72.9.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berg D. E., Egner C., Hirschel B. J., Howard J., Johnsrud L., Jorgensen R. A., Tlsty T. D. Insertion, excision, and inversion of Tn5. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):115–123. doi: 10.1101/sqb.1981.045.01.020. [DOI] [PubMed] [Google Scholar]
- Beringer J. E. R factor transfer in Rhizobium leguminosarum. J Gen Microbiol. 1974 Sep;84(1):188–198. doi: 10.1099/00221287-84-1-188. [DOI] [PubMed] [Google Scholar]
- Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
- Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guiney D. G., Helinski D. R. The DNA-protein relaxation complex of the plasmid RK2: location of the site-specific nick in the region of the proposed origin of transfer. Mol Gen Genet. 1979 Oct 3;176(2):183–189. doi: 10.1007/BF00273212. [DOI] [PubMed] [Google Scholar]
- Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
- Johnson R. C., Reznikoff W. S. Localization of the Tn5 transposase promoter using the cycling reaction of RNA polymerase. Nucleic Acids Res. 1981 Apr 24;9(8):1873–1883. doi: 10.1093/nar/9.8.1873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson R. C., Yin J. C., Reznikoff W. S. Control of Tn5 transposition in Escherichia coli is mediated by protein from the right repeat. Cell. 1982 Oct;30(3):873–882. doi: 10.1016/0092-8674(82)90292-6. [DOI] [PubMed] [Google Scholar]
- Jorgensen R. A., Rothstein S. J., Reznikoff W. S. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. doi: 10.1007/BF00267254. [DOI] [PubMed] [Google Scholar]
- Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
- Matsuhashi Y., Sawa T., Takeuchi T., Umezawa H. Purification of aminoglycoside 3'-phosphotransferase II. J Antibiot (Tokyo) 1976 Feb;29(2):204–207. doi: 10.7164/antibiotics.29.204. [DOI] [PubMed] [Google Scholar]
- Meade H. M., Long S. R., Ruvkun G. B., Brown S. E., Ausubel F. M. Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol. 1982 Jan;149(1):114–122. doi: 10.1128/jb.149.1.114-122.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meade H. M., Signer E. R. Genetic mapping of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1977 May;74(5):2076–2078. doi: 10.1073/pnas.74.5.2076. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray K., Murray N. E. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli. J Mol Biol. 1975 Nov 5;98(3):551–564. doi: 10.1016/s0022-2836(75)80086-6. [DOI] [PubMed] [Google Scholar]
- Putnoky P., Kiss G. B., Ott I., Kondorosi A. Tn5 carries a streptomycin resistance determinant downstream from the kanamycin resistance gene. Mol Gen Genet. 1983;191(2):288–294. doi: 10.1007/BF00334828. [DOI] [PubMed] [Google Scholar]
- Roberts R. J. Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res. 1983 Jan 11;11(1):r135–r167. [PMC free article] [PubMed] [Google Scholar]
- Rothstein S. J., Jorgensen R. A., Postle K., Reznikoff W. S. The inverted repeats of Tn5 are functionally different. Cell. 1980 Mar;19(3):795–805. doi: 10.1016/s0092-8674(80)80055-9. [DOI] [PubMed] [Google Scholar]
- Rothstein S. J., Reznikoff W. S. The functional differences in the inverted repeats of Tn5 are caused by a single base pair nonhomology. Cell. 1981 Jan;23(1):191–199. doi: 10.1016/0092-8674(81)90284-1. [DOI] [PubMed] [Google Scholar]
- Selvaraj G., Iyer V. N. A dnaB analog function specified by bacteriophage P7 and its comparison to the similar function specified by bacteriophage P1. Mol Gen Genet. 1980;178(3):561–566. doi: 10.1007/BF00337861. [DOI] [PubMed] [Google Scholar]
- Selvaraj G., Iyer V. N. Genetic transformation of Rhizobium meliloti by plasmid DNA. Gene. 1981 Nov;15(2-3):279–283. doi: 10.1016/0378-1119(81)90137-2. [DOI] [PubMed] [Google Scholar]
- Selvaraj G., Iyer V. N. Suicide plasmid vehicles for insertion mutagenesis in Rhizobium meliloti and related bacteria. J Bacteriol. 1983 Dec;156(3):1292–1300. doi: 10.1128/jb.156.3.1292-1300.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shaw K. J., Berg C. M. Escherichia coli K-12 auxotrophs induced by insertion of the transposable element Tn5. Genetics. 1979 Jul;92(3):741–747. doi: 10.1093/genetics/92.3.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanaka T., Weisblum B. Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J Bacteriol. 1975 Jan;121(1):354–362. doi: 10.1128/jb.121.1.354-362.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thatte V., Iyer V. N. Cloning of a plasmid region specifying the N transfer system of bacterial conjugation in Escherichia coli. Gene. 1983 Mar;21(3):227–236. doi: 10.1016/0378-1119(83)90006-9. [DOI] [PubMed] [Google Scholar]
- Umezawa H. Biochemical mechanism of resistance to aminoglycosidic antibiotics. Adv Carbohydr Chem Biochem. 1974;30:183–225. doi: 10.1016/s0065-2318(08)60265-6. [DOI] [PubMed] [Google Scholar]
- Wood W. B. Host specificity of DNA produced by Escherichia coli: bacterial mutations affecting the restriction and modification of DNA. J Mol Biol. 1966 Mar;16(1):118–133. doi: 10.1016/s0022-2836(66)80267-x. [DOI] [PubMed] [Google Scholar]