Summary
A lambda transducing phage carrying the traGSTD genes of the E. coli K12 sex factor F was isolated by an in vivo technique, and characterized in ra complementation tests, by determining its restriction endonuclease fragment sizes, and by measuring heteroduplex molecules. The size and location on the F physical map of the tra transducing segment was thereby determined.
Comparison of the proteins synthesized in UV-irradiated cells by this phage and by a derivative carrying the amber traG79 mutation, allowed the traG product to be identified as a protein of molecular weight 100,000. In the same experiments, the sizes of the traT and traD products made by the phage were also measured, being 25,000 and 85,000 daltons respectively.
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Achtman, M., Kennedy, N., Helmuth, R., Morelli, G., Beutin, L., Thompson, R.: Cell-cell interactions in conjugating E. coli: conjugation proteins and the mating cycle. In: Microbial drug resistance (S. Mitsuhashi, ed.), Vol. 2, pp. 157–168. Tokyo: Pub. Japan Scientific Societies Press 1979
Achtman, M., Skurray, R.: A redefinition of the mating phenomenon in bacteria. In: Microbial interactions (J.L. Reissig, ed.), Vol. 3, pp. 232–279. London: Pub. Chapman and Hall 1977
Achtman, M., Skurray, R.A., Thompson, R., Helmuth, R., Hall, S., Beutin, L., Clark, A.J.: Assignment of tra cistrons to EcoRI fragments of F sex factor DNA. J. Bacteriol. 133, 1383–1392 (1978)
Achtman, M., Willetts, N.S., Clark, A.J.: Beginning a genetic analysis of conjugational transfer determined by the F factor in E. coli by isolation and characterisation of transfer-deficient mutants. J. Bacteriol. 106, 529–538 (1971)
Achtman, M., Willetts, N.S., Clark, A.J.: A conjugational complementation analysis of transfer-deficient mutants of Flac in E. coli. J. Bacteriol. 110, 831–842 (1972)
Blattner, F.R., Fiandt, M., Hass, K.K., Twose, P.A., Szybalski, W.: Deletions and insertions in the immunity region of coliphage lambda: revised measurements of the promoter-startpoint distance. Virology 62, 458–471 (1974)
Clark, A.J., Skurray, R.A., Crisona, N.J., Nagaishi, H.: Use of molecular cloning in the genetic and functional analysis of the plasmid F. In: Molecular mechanisms in the control of gene expression (D.P. Nierlich, W.J. Rutter and C.F. Fox, eds.), pp. 565–579. New York: Pub. Academic Press Inc. 1976
Enquist, L.W., Weisberg, R.A.: The red plaque test: a rapid method for identification of excision defective variants of bacteriophage lambda. Virol. 72, 147–153 (1976)
Everett, R., Willetts, N.S.: Characterisation ofan in vivo system for nicking at the origin of conjugal DNA transfer of the sex factor F. J. Mol. Biol., 136, 129–150 (1980)
Haggerty, D.M., Schleif, R.F.: Location in bacteriophage lambda DNA of cleavage sites of the site-specific endonuclease from B. amyloliquefaciens H. J. Virol. 18, 659–663 (1976)
Ippen-Ihler, K., Achtman, M., Willetts, N.S.: A deletion map of the E. coli K12 sex factor F: the order of eleven transfer cistrons. J. Bacteriol. 110, 857–863 (1972)
Kennedy, M., Beutin, L., Achtman, M., Skurray, R., Rahmsdorf, U., Herrlich, P.: Conjugation proteins encoded by the F sex factor. Nature 270, 580–585 (1977)
Kingsman, A., Willetts, N.: The requirements for conjugal DNA synthesis in the donor strain during Flac transfer. J. Mol. Biol. 122, 287–300 (1978)
Landy, A., Ross, W.: Viral integration and excision: structure of the lambda att sites. Science 197, 1147–1160 (1977)
Linn, T., Goman, M., Scaife, J.G.: Studies on the control of the genes for transcription and translation in E. coli K12. I. tufB and rplA,K have separate promoters. J. Mol. Biol. 130, 405–420 (1979)
McIntire, S., Willetts, N.S.: Transfer-deficient cointegrates of Flac and lambda prophage. Mol. Gen. Genet. 178, 165–172 (1980)
McParland, R.H., Brown, L.R., Pearson, G.D.: Cleavage of lambda DNA by a site-specific nuclease from Serratia marcescens. J. Virol. 19, 1006–1011 (1976)
Miki, T., Horiuchi, T., Willetts, N.S.: Identification and characterisation of four new tra cistrons on the E. coli sex factor F. Plasmid 1, 316–323 (1978)
Novotny, C., Fives-Taylor, P.: Retraction of F pili J. Bacteriol. 177, 1306–1311 (1974)
Ohtsubo, E., Nishimura, Y., Hirota, Y.: Transfer-defective mutants of sex factors in E. coli. I. Defective mutants and complementation analysis. Genetics 64, 173–188 (1970)
Ohtsubo, H., Ohtsubo, E.: Repeated DNA sequences in plasmids, phages, and bacterial chromosomes. In: DNA insertion elements plasmids and episomes (A.I. Bukhari, J.A. Shapiro, and S. Adhya, eds.), pp. 49–64. New York: Cold Spring Harbor Laboratory 1977
Philippsen, P., Kramer, R.A., Davis, R.W.: Cloning of the yeast ribosomal DNA repeat unit in SstI and HindIII lambda vectors using genetic and physical size selections. J. Mol. Biol. 123, 371–386 (1978)
Willetts, N.S.: Mapping loci for surface exclusion and incompatibility on the F factor of E. coli K12. J. Bacteriol. 118, 778–782 (1974)
Willetts, N.S.: Genetics of conjugation. In: R factor-drug resistance plasmid (S. Mitsuhashi, ed), pp 89–107. Tokyo: University of Tokyo Press 1977
Willetts, N., Johnson, G.: R. SmaI cleavage map of the transfer region of the E. coli K12 sex factor F. Genet. Res. 34, 195–202 (1979)
Willetts, N., Maule, J.: Investigations of the F conjugation gene traI: traI mutants and λtraI transducing phages. Mol. Gen. Genet. 169, 325–336 (1979)
Willetts, N.S., McIntire, S.: Isolation and characterisation of λtra transducing phages from EDFL223 (Flac traB::EDλ4). J. Mol. Biol. 126, 525–549 (1978)
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Willetts, N., Maule, J. Characterisation of a λ transducing phage carrying the F conjugation gene traG . Molec. gen. Genet. 178, 675–680 (1980). https://doi.org/10.1007/BF00337878
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DOI: https://doi.org/10.1007/BF00337878