Abstract
A Mu d1 derivative is described which is useful for genetic manipulation of Mu-lac fusion insertions. A double mutant of the specialized transducing phage Mu d1(Amp Lac c62ts) was isolated which is conditionally defective in transposition ability. The Mu d1 derivative, designated Mu d1-8(Tpn[Am] Amp Lac c62ts), carries mutations which virtually eliminate transposition in strains lacking an amber suppressor. In such strains, the Mu d1-8 prophage behaves like a standard transposon. It can be moved from one strain of Salmonella typhimurium to another by the general transducing phage P22 with almost 100% inheritance of the donor insertion mutation. When introduced into a recipient carrying supD, supE, or supF, 89 to 94% of the Ampr transductants were transpositions of the donor Mu d1-8, from the transduced fragment into new sites. The stability of Mu d1-8 in a wild-type, suppressor-free background was sufficient to permit use of the fusion to select constitutive mutations without prior isolation of deletions to stabilize the fusion. Fusion strains could be grown at elevated temperature without induction of the Mu d prophage. The transposition defect of Mu d1-8 was corrected by a plasmid carrying the Mu A and B genes.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson P., Roth J. Spontaneous tandem genetic duplications in Salmonella typhimurium arise by unequal recombination between rRNA (rrn) cistrons. Proc Natl Acad Sci U S A. 1981 May;78(5):3113–3117. doi: 10.1073/pnas.78.5.3113. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baker T. A., Howe M. M., Gross C. A. Mu dX, a derivative of Mu d1 (lac Apr) which makes stable lacZ fusions at high temperature. J Bacteriol. 1983 Nov;156(2):970–974. doi: 10.1128/jb.156.2.970-974.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beckwith J. A genetic approach to characterizing complex promoters in E. coli. Cell. 1981 Feb;23(2):307–308. doi: 10.1016/0092-8674(81)90125-2. [DOI] [PubMed] [Google Scholar]
- Beny G., Boyen A., Charlier D., Lissens W., Feller A., Glansdorff N. Promoter mapping and selection of operator mutants by using insertion of bacteriophage Mu in the argECBH divergent operon of Escherichia coli K-12. J Bacteriol. 1982 Jul;151(1):62–67. doi: 10.1128/jb.151.1.62-67.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukhari A. I. Reversal of mutator phage Mu integration. J Mol Biol. 1975 Jul 25;96(1):87–99. doi: 10.1016/0022-2836(75)90183-7. [DOI] [PubMed] [Google Scholar]
- Casadaban M. J., Chou J. In vivo formation of gene fusions encoding hybrid beta-galactosidase proteins in one step with a transposable Mu-lac transducing phage. Proc Natl Acad Sci U S A. 1984 Jan;81(2):535–539. doi: 10.1073/pnas.81.2.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
- Chan R. K., Botstein D., Watanabe T., Ogata Y. Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high-frequency-transducing lysate. Virology. 1972 Dec;50(3):883–898. doi: 10.1016/0042-6822(72)90442-4. [DOI] [PubMed] [Google Scholar]
- Demerec M., Adelberg E. A., Clark A. J., Hartman P. E. A proposal for a uniform nomenclature in bacterial genetics. Genetics. 1966 Jul;54(1):61–76. doi: 10.1093/genetics/54.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giphart-Gassler M., Goosen T., van Meeteren A., Wijffelman C., van de Putte P. Properties of the recombinant plasmid pGP1 containing part of the early region of bacteriophage mu. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1179–1185. doi: 10.1101/sqb.1979.043.01.133. [DOI] [PubMed] [Google Scholar]
- Giphart-Gassler M., Van de Putte P. Thermo-inducible expression of cloned early genes of bacteriophage Mu. Gene. 1979 Sep;7(1):33–50. doi: 10.1016/0378-1119(79)90041-6. [DOI] [PubMed] [Google Scholar]
- Holley E. A., Foster J. W. Bacteriophage P22 as a vector for Mu mutagenesis in Salmonella typhimurium: isolation of nad-lac and pnc-lac gene fusions. J Bacteriol. 1982 Nov;152(2):959–962. doi: 10.1128/jb.152.2.959-962.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong J. S., Ames B. N. Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3158–3162. doi: 10.1073/pnas.68.12.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hughes K. T., Cookson B. T., Ladika D., Olivera B. M., Roth J. R. 6-Aminonicotinamide-resistant mutants of Salmonella typhimurium. J Bacteriol. 1983 Jun;154(3):1126–1136. doi: 10.1128/jb.154.3.1126-1136.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kenyon C. J., Walker G. C. DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci U S A. 1980 May;77(5):2819–2823. doi: 10.1073/pnas.77.5.2819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Komeda Y., Iino T. Regulation of expression of the flagellin gene (hag) in Escherichia coli K-12: analysis of hag-lac gene fusions. J Bacteriol. 1979 Sep;139(3):721–729. doi: 10.1128/jb.139.3.721-729.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maloy S. R., Roth J. R. Regulation of proline utilization in Salmonella typhimurium: characterization of put::Mu d(Ap, lac) operon fusions. J Bacteriol. 1983 May;154(2):561–568. doi: 10.1128/jb.154.2.561-568.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mulligan J. T., Margolin W., Krueger J. H., Walker G. C. Mutations affecting regulation of methionine biosynthetic genes isolated by use of met-lac fusions. J Bacteriol. 1982 Aug;151(2):609–619. doi: 10.1128/jb.151.2.609-619.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray M. L., Hartman P. E. Overproduction of hisH and hisF gene products leads to inhibition of cell cell division in Salmonella. Can J Microbiol. 1972 May;18(5):671–681. doi: 10.1139/m72-105. [DOI] [PubMed] [Google Scholar]
- Ornellas E. P., Stocker B. A. Relation of lipopolysaccharide character to P1 sensitivity in Salmonella typhimurium. Virology. 1974 Aug;60(2):491–502. doi: 10.1016/0042-6822(74)90343-2. [DOI] [PubMed] [Google Scholar]
- Ratzkin B., Roth J. Cluster of genes controlling proline degradation in Salmonella typhimurium. J Bacteriol. 1978 Feb;133(2):744–754. doi: 10.1128/jb.133.2.744-754.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenfeld S. A., Brenchley J. E. Bacteriophage P1 as a vehicle for Mu mutagenesis of Salmonella typhimurium. J Bacteriol. 1980 Nov;144(2):848–851. doi: 10.1128/jb.144.2.848-851.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roth J. R., Antón D. N., Hartman P. E. Histidine regulatory mutants in Salmonella typhimurium. I. Isolation and general properties. J Mol Biol. 1966 Dec 28;22(2):305–323. doi: 10.1016/0022-2836(66)90134-3. [DOI] [PubMed] [Google Scholar]
- Silhavy T. J., Benson S. A., Emr S. D. Mechanisms of protein localization. Microbiol Rev. 1983 Sep;47(3):313–344. doi: 10.1128/mr.47.3.313-344.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
- Van Leerdam E., Karreman C., van de Putte P. Ner, a cro-like function of bacteriophage Mu. Virology. 1982 Nov;123(1):19–28. doi: 10.1016/0042-6822(82)90291-4. [DOI] [PubMed] [Google Scholar]
- Waggoner B., Pato M., Toussaint A., Faelen M. Replication of mini-Mu prophage DNA. Virology. 1981 Aug;113(1):379–387. doi: 10.1016/0042-6822(81)90163-x. [DOI] [PubMed] [Google Scholar]
- Wanner B. L., McSharry R. Phosphate-controlled gene expression in Escherichia coli K12 using Mudl-directed lacZ fusions. J Mol Biol. 1982 Jul 5;158(3):347–363. doi: 10.1016/0022-2836(82)90202-9. [DOI] [PubMed] [Google Scholar]
- Wanner B. L., Wieder S., McSharry R. Use of bacteriophage transposon Mu d1 to determine the orientation for three proC-linked phosphate-starvation-inducible (psi) genes in Escherichia coli K-12. J Bacteriol. 1981 Apr;146(1):93–101. doi: 10.1128/jb.146.1.93-101.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Worsham P. L., Konisky J. Use of cir-lac operon fusions to study transcriptional regulation of the colicin Ia receptor in Escherichia coli K-12. J Bacteriol. 1981 Jan;145(1):647–650. doi: 10.1128/jb.145.1.647-650.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]