Summary
Twelve transfer-deficient mutants of the plasmid Flac were obtained by insertion of prophage lambda into secondary attachment sites within the transfer region. Insertions into eight different tra genes were identified. These mutations were strongly polar on expression of tra genes previously mapped “downstream”, and thus confirmed that the genes traA through traD form a single operon. However, some continued expression of traI suggested that this was transcribed in part from a promoter located between traD and tral, and in part from the transfer operon promoter. One insertion early in the transfer operon produced a plasmid-specific tra mutation not complemented by R100-1 or R1-19: this insertion was into a new gene (traY), located before traA as the first member of the transfer operon. Partial tra deletion mutants were obtained as 42° C — survivors from several of the Flac tra:: EDλ4 plasmids, and their properties are described.
Similar content being viewed by others
References
Achtman, M., Kennedy, N., Skurray, R.: Cell-cell interactions in conjugating Escherichia coli: role of traT protein in surface exclusion. Proc. Natl. Acad. Sci. U.S.A. 74, 5104–5108 (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 Escherichia coli by isolation and characterization of transferdeficient mutants. J. Bacteriol. 106, 529–538 (1971a)
Achtman, M., Willetts, N.S., Clark, A.J.: A conjugational complementation analysis of transfer-deficient mutants of Flac in Escherichia coli. J. Bacteriol. 110, 831–842 (1971b)
Cooke, M., Meynell, E.: Chromosomal transfer mediated by derepressed R factors in F- Escherichia coli K12. Genet. Res. 14, 79–87 (1969)
Dempsey, W.B., Willetts, N.S.: Plasmid cointegrates of prophage lambda and R factor R100. J. Bacteriol. 126, 166–176 (1976)
Egawa, R., Hirota, Y.: Inhibition of fertility by multiple drugresistance factor in Escherichia coli K12. Japan. J. Genet. 37, 66–69 (1962)
Everett, R., Willetts, N.: Characterization of an in vivo system for nicking at the origin of conjugational DNA transfer of the sex factor F. J. Mol. Biol. (in press) (1979)
Foster, T.J., Willetts, N.S.: Genetic analysis of deletions of R100-1 that are both transfer-deficient and tetracycline-sensitive. J. Gen. Microbiol. 93, 133–140 (1976)
Foster, T.J., Willetts, N.S.: Characterization of transfer-deficient mutants of the R100-1 Tcs plasmid pDU202, caused by insertion of Tn10. Mol. Gen. Genet. 156, 107–114 (1977)
Helmuth, R., Achtman, M.: Operon structure of DNA transfer cistrons on the F sex factor. Nature 257, 652–656 (1975)
Ippen-Ihler, K., Achtman, M., Willetts, N.S.: A deletion map of the Escherichia coli K12 sex factor F: the order of eleven transfer cistrons. J. Bacteriol. 110, 857–863 (1972)
Low, K.B.: Escherichia coli K12 F-prime factors, old and new. Bacteriol. Rev. 36, 587–607 (1973)
McIntire, S., Willetts, N.S.: Plasmid cointegrates of Flac and lambda prophage. J. Bacteriol. 134, 184–192 (1978)
Miki, T., Horiuchi, T., Willetts, N.S.: Identification and characterization of four new tra cistrons on the Escherichia coli sex factor F. Plasmid 1, 316–323 (1978)
Ohtsubo, E., Nishimura, Y., Hirota, Y.: Transfer-defective mutants of sex factors in Escherichia coli I. Defective mutants and complementation analysis. Genetics 64, 173–188 (1970)
Shimada, K., Campbell, A.: Int-constitutive mutants of bacteriophage lambda. Proc. Natl. Acad. Sci. U.S.A. 71, 237–241 (1974)
Shimada, K., Weisberg, R.A., Gottesman, M.E.: Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J. Mol. Biol. 63, 483–503 (1972)
Thomspon, R., Achtman, M.: The control region of the F sex factor DNA transfer cistrons: physical mapping by deletion analysis. Mol. Gen. Genet. 169, 49–57 (1979)
Willetts, N.S.: The plasmid-specificity of two proteins required for conjugation in Escherichia coli K12. Nature New Biol 230, 183–185 (1971)
Willetts, N.S.: Characterization of the F transfer cistron, traL. Genet. Res. 21, 205–213 (1973)
Willetts, N.S.: Mapping loci for surface exclusion and incompatibility on the F factor of Escherichia coli K12. J. Bacteriol. 118, 778–782 (1974)
Willetts, N.S.: The transcriptional control of fertility in F-like plasmids. J. Mol. Biol. 112, 141–148 (1977a)
Willetts, N.S.: Genetics of conjugation. In: R factor-drug resisance plasmid (S. Mitsuhashi, ed.), pp. 89–107. Tokyo: Univ. Tokyo Press 1977b
Willetts, N.S.: Control of conjugation in F-like plasmids. In: Microbiology — 1978 (D. Schlessinger, ed.), pp. 211–213, Washington, D.C.: American Society for Microbiology 1978
Willetts, N.S., Achtman, M.: A genetic analysis of trasfer by the Escherichia coli sex factor F, using P1 transductional complementation. J. Bacteriol 110, 843–851 (1972)
Willetts, N.S., Maule, J.: Investigations of the F conjugation gene traI: traI mutants and λraI transducing phages. Mol. Gen. Genet. 169, 325–336 (1979)
Willetts, N.S., McIntire, S.: Isolation and characterization of λtra transducing phages from EDFL223 (Flac traB::EDλ4). J. Mol. Biol. 126, 525–549 (1978)
Author information
Authors and Affiliations
Additional information
Communicated by G.O'Donovan
Rights and permissions
About this article
Cite this article
McIntire, S., Willetts, N. Transfer-deficient cointegrates of Flac and lambda prophage. Molec. Gen. Genet. 178, 165–172 (1980). https://doi.org/10.1007/BF00267225
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267225