Abstract
Erwinia chrysanthemi CUCPB 1237 produces multiple isozymes of pectate lyase (PL). Marker exchange mutagenesis of the pelB gene was accomplished with a cloned pe1B gene that had been inactivated in vitro by insertion of a kanamycin resistance determinant. The E. chrysanthemi mutant was Kanr, Amps, lacked pBR322 sequences, and was deficient only in PLb, as determined by isoelectric focusing. Although PLb is one of the four major PL isozymes produced by this strain, the PLb-deficient mutant differed little from the wild type parent in growth rate or overall PL production in a medium containing polygalacturonic acid as the sole carbon source. Nor was any difference detected in the ability of the mutant to cause maceration in intact potato tubers.
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References
Collmer A and Bateman DF. 1981. Impaired induction and self-catabolite repression of extra cellular pectate lyase in Erwinia chrysanthemi mutants deficient in oligogalacturonide lyase. Proc. Natl. Acad. Sci. U.S.A. 78:3920–3924.
Collmer A, Schoedel C, Roeder DL, Ried JL, and Rissler JF. 1985. Molecular cloning in Escherichia coli of Erwinia chrysanthemi genes encoding multiple forms of pectate lyase. J. Bacteriol. 161:913–920.
Keen NT, Dahlbeck D, Staskawicz B, and Belser W. 1984. Molecular cloning of pectate lyase genes from Erwinia chrysanthemi and their expression in Escherichia coli. J. Bacteriol. 159:825–831.
Kotoujansky A, Diolez A, Boccara B, Bertheau Y, Andro T, and Coleno A. 1985. Molecular cloning of Erwinia chrysanthemi pectinase and cellulase structural genes. EMBO J. 4:781–785.
Leary JL, Brigati DF, and Ward DC. 1983. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: bio-blots. Proc. Natl. Acad. Sci. USA 80:4054–4049.
Pupillo P, Mazzucchi U, and Pierini G. 1976. Pectic lyase isozyme produced by Erwinia chrysanthemi Burkh. et al. in polypectate broth or in Dieffenbachi leaves. Physiol. Plant Pathol. 9:113–120.
Ried JL and Collmer A. 1985. An activity stain for the rapid characterization of pectic enzymes in isoelectric focusing and sodium dodecyl sulfate polyacrylamide gels. Appl. Envir. Microbiol. 50:(in press).
Roeder DL and Collmer A. 1985. Marker-exchange mutagenesis of a pectate lyase isozyme gene in Erwinia chrysanthemi. J. Bacteriol. 164:(in press).
Southern E. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98:503–517.
Van Haute E, Joos H, Maes M, Warren G, Van Montagu M, and Schell, J. 1983. Intergeneric transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti plasmids of Agrobacterium tumefaciens. EMBO J. 2:411–417.
Van Gijsaegem F, Toussaint A, and Schoonagana E. 1985. In vivo cloning of the pectate lyase and cellulase genes of Erwinia chrysanthemi. EMBO J. 4:787–792.
Vieira J and Messing J. 1982. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268.
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© 1987 Martinus Nijhoff Publishers, Dordrecht
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Roeder, D.L., Collmer, A. (1987). Marker—Exchange Mutagenesis of the pe1B Gene in Erwinia Chrysanthemi CUCPB 1237. In: Civerolo, E.L., Collmer, A., Davis, R.E., Gillaspie, A.G. (eds) Plant Pathogenic Bacteria. Current Plant Science and Biotechnology in Agriculture, vol 4. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-3555-6_34
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DOI: https://doi.org/10.1007/978-94-009-3555-6_34
Publisher Name: Springer, Dordrecht
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