Summary
Phage Mud1 cts (Apr lac), or Mud1, insertion mutations may be accompanied by adjacent deletion formation which can complicate use of lac fusions generated with this phage for gene regulatory studies. As for phage Mu insertion mutations, phage Mud1 insertions fail to revert at significant frequency (whether or not accompanied by an adjacent deletion). We describe isolation of revertible (X mutant) derivatives of phage Mud1 in Salmonella typhimurium. The X mutant derivatives allow use of reversion as a simple test to determine whether a Mud1 insertion has occurred precisely without an adjacent deletion that may have fused the lac genes to a promoter outside of the gene of interest. In addition, a simple method for stabilizing Mud1 generated lac fusions against subsequent transposition is described.
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References
Alper MD, Ames BN (1978) Transport of antibiotics and metabolite analogs by systems under cyclic AMP control: positive selection of Salmonella typhimurium cya and crp mutants. J Bacteriol 133:149–157
Baker TA, Howe MM, Gross CA (1983) Mu dX, a derivative of Mu d1 (lac Apr) which makes stable lacZ fusions at high temperature. J Bacteriol 156:970–974
Bankaitis VA, Bassford Jr PJ (1982) Regulation of adenylate cyclase synthesis in Escherichia coli: studies with cya-lac operon and protein fusion strains. J Bacteriol 151:1346–1357
Berman ML, Beckwith J (1979) Fusion of the lac operon to the transfer RNA gene tyrT of Escherichia coli. J Mol Biol 130:285–301
Blum P, Blaha L, Shand R, Holzschu D, Artz S (1982) Manipulations of phage Mud1 (lac Apr) in Salmonella typhimurium and use of a his-lac fusion to select his promoter mutants. Am Soc Microbiol Abs H 128–134
Bukhari AI (1975) Reversal of mutator phage Mu integration. J Mol Biol 96:87–99
Bukhari AI, Froshauer S (1978) Insertion of a transposon for chloramphenicol resistance into bacteriophage Mu. Gene 3:303–314
Cabezon T, Faelen M, DeWilde M, Bollen A, Thomas R (1975) Expression of ribosomal protein genes in Escherichia coli. Mol Gen Genet 137:125–129
Casadaban MJ, Cohen SN (1979) 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 USA 76:4530–4533
Csonka LN, Howe MM, Ingraham JL, Pierson III LS, Turnbough Jr CL (1981) Infection of Salmonella typhimurium with coliphage Mud1 (Apr lac): construction of pyr::lac gene fusions. J Bacteriol 145:299–305
Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Dixon R, Eady RR, Espin G, Hill S, Iaccarino M, Kahn D, Merrick M (1980) Analysis of regulation of Klebsiella pneumonia nitrogen fixation (nif) gene cluster with gene fusions. Nature 286:128–132
Enomoto M, Stocker BAD (1974) Transduction by phage P1kc in Salmonella typhimurium. Virology 60:503–514
Gigot D, Crabeel M, Feller A, Charlier D, Lissens W, Glansdorff N, Pierard A (1980) Patterns of polarity in the Escherichia coli carAB gene cluster. J Bacteriol 143:914–920
Howe MM, Zipser D (1974) Host deletions caused by the integration of bacteriophage Mu-1. Am Soc Microbiol Abs V 208:235
Hughes KT, Roth JR (1984) Conditionally transposition-defective derivative of Mu d1 (Amp Lac). J Bacteriol 159:130–137
Jovanovich SB (1985) Regulation of a cya-lac fusion by cyclic AMP in Salmonella typhimurium. J Baceriol 161:641–649
Jovanovich SB, Couper L, Artz S (1983) Regulation of a cya-lac fusion in Salmonella typhimurium. Am Soc Microbiol Abs H 2:106
Lee JH, Heffernan L, Wilcox G (1980) Isolation of ara-lac gene fusion in Salmonella typhimurium LT2 by using transducing bacteriophage Mu d (Apr lac). J Bacteriol 143:1325–1331
Lee JH, Al-Zarban S, Wilcox G (1981) Genetic characterization of the araE gene in Salmonella typhimurium LT2. J Bacteriol 146:298–304
Maloy SR, Roth JR (1983) Regulation of proline utilization in Salmonella typhimurium: characterization of put::Mu d(Ap, lac) operon fusions. J Bacteriol 154:561–568
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
O'Connor MBI, Malamy MH (1983) A new insertion sequence, IS121, is found on the Mu d1 (Ap lac) bacteriophage and the Escherichia coli chromosome. J Bacteriol 156:669–679
Rosenfeld SA, Brenchley JE (1980) Bacteriophage P1 as a vehicle for Mu mutagenesis of Salmonella typhimurium. J Bacteriol 144:848–851
Roy A, Haziza C, Danchin A (1983) Regulation of adenylate cyclase synthesis in Escherichia coli: nucleotide sequence of the control region. EMBO J 2:791–797
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Communicated by M.M. Green
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Blum, P., Blaha, L. & Artz, S. Reversion and immobilization of phage Mud1 cts (Apr lac) insertion mutations in Salmonella typhimurium . Mol Gen Genet 202, 327–330 (1986). https://doi.org/10.1007/BF00331659
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DOI: https://doi.org/10.1007/BF00331659