Full text in pdf format

Cloning of structural genes for colicin V and their role in pathogenicity of invasive Escherichia coli

Marilda Carlos Vidotto^I; Célia Regina Goes^I; Júlia Taque^I; Amilcar Tanuri^I; Diógenes Santiago Santos^II

^IDepartamento de Patologia Geral, CCB, Universidade Estadual de Londrina, 86051 Londrina, PR, Brasil. Send correspondence to M.C.V.
^IICentro dc Biotecnologia, Universidade Federal do Rio Grande do Sul, 90000 Porto Alegre, RS, Brasil

ABSTRACT

The colicin V structural genes of plasmid pMV₁₄ were cloned into he vector pYP 328. These genes were isolated on a 2.4 kb deoxyribonucleic acid fragment which did not transport or express the virulence genes of strain UEL 14. Insertion of the transposon Tn5 into plasmid pMV14 led to the construction of a mutant plasmid which did not produce colicin V but conferred virulence to one-day-old chicks. From these results we conclude that the colicin V activity is not essential to ColV plasmid-mediated virulence enhancement in Escherichia coil of avian origin.

Keywords: Colicin V; Pathogenicity; Escherichia coli.

REFERENCES

Azevedo, J.L. and Costa, S.O.P. (1973). Exercícios Práticos de Genética. Edit. Nacional e Edusp, Sao Paulo.

Birnboin, H.C. and Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic. Acids. Res. 7: 1513.

Boyer, H.W. and Roulland-Doussoix, D. (1%9). Complementation analysis of the restriction and modification of DNA in E. coli. J. Mol. Biol. 41: 459-472.

Falkow, S., Guerry, Hedges, R.W. and Datta, N. (1974). Polynucleotide sequence relationships among plasmids of the I compatibility complex. J. Microbiol. 85: 65-76.

Frick, KK, Quakenbush, R.L. and Konisky, J. (1981). Cloning of immunity and structural genes for colicin V. J. Bacterial 148: 498-507.

Gilson, L., Mahanty, H.K. and Kolter, R. (1987). Four plasmid genes are required for colicin V synthesis, export and immunity. J. Bacteriol. 169: 2466-2470.

Hardy, KG. (1975). Colicinogeny and related phenomena. Bacteriol. Rev. 39: 464-515.

Harry, E.G. (1964). A study of 119 outbreaks of coli-septicaemia in broiler flocks. Et. Rec. 76: 443-449.

Kado, C.I. and Liu, S.T. (1981). Rapid procedure for the detection and isolation of large and small plasmids. J. Bacteriol. 145: 1365-1373.

Maniatis, T., Fritsch, E.F. and Sambrook, J. (1982). Molecular cloning: a laboratory manual. Cold. Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

Meyers, J.A., Sanches, D., Elnell, L.D. and Falkow, S. (1976). Simple agarose gel electrophoretic method for the identification and characterization of Plasmid Deoxyribonucleic Acid. J. Bacteriol. 127: 1529-1537.

Milch, H., Nikolnikov, S. and Cziróx, E. (1984). Escherichia coli Col V plasmids and their role in pathogenicity. Acta Microbiol. Hung. 31: 117-125.

Ozanne, G., Mathieu, L.G. and Baril, J.P. (1977). Production of Colicin V. In vitro and in vivo and observations on its effect in experimental animals. Infect. Immun. 17: 497-503.

Quackensbush, R.L. and Falkow, S. (1979). Relationship between colicin V activity and virulence in E. coll. Infect. Immun. 24: 562-564.

Silva, M.L.M., Maas, W.K. and Gyles, C.L. (1978). Isolation and characterization of enterotoxin - deficient mutants of E. coli. Proc. Nalt. Acad. Sci. USA 75; 1384-1388.

Smith, H.W. (1974). A search for transmissible pathogenic characters, in invasive strains of Escherichia coli the discovery of a plasmid-controlled toxin and a plasmid-controled lethal character closely associated, or identical, with colicin V.J. Gen. Microbiol. 83: 95-111.

Smith, H.W. and Huggins, M.B. (1976). Further observations on the colicin V plasmid of Escherichia coli with pathogenicity and with survival in the alimentary tract. J. Gen. Microbiol. 92: 335-350.

Teixeira, S.M.R. (1985). Clonagem de seqüências de replicação autônomas (ARS) de Trypanosoma cruzi. Masters-Thesis, Instituto de Ciências Biológicas da Universidade de Brasilia, Brasília, DF.

Vidotto, M.C. (1988). Caracterização e estudo genético-molecular de fatores de virulência de Escherichia coli aviária. Doctoral Thesis, Escola Paulista de Medicina, São Paulo, SP.

Vidotto, M.C., Guimarães, L.G., Müller, E.E., Freitas, J.C., Alfieri, A.A. and Santos, D.S. (1990). Virulence factors of avian Escherichia coli. Avian Diseases 34: 531-538.

Yang, C.C. and Konisky, J. (1984). Colicin V treated Escherichia coli does not generate membrane potential. J. Bacteriol. 158: 757-759.