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. 1985 Sep;163(3):913–917. doi: 10.1128/jb.163.3.913-917.1985

Mu-lac insertion-directed mutagenesis in a pectate lyase gene of Erwinia chrysanthemi.

A Diolez, A Coleno
PMCID: PMC219219  PMID: 2993251

Abstract

The pelC gene, which encodes one of the five major pectate lyase (PL) isoenzymes in Erwinia chrysanthemi 3937, designated PLc, was subcloned from a hybrid lambda phage into a pBR322 derivative and mutagenized with a mini-Mu-lacZ transposable element able to form fusions to the lacZ gene. One plasmid (pAD1) which had an inactivated pelC gene and a Lac+ phenotype was selected in Escherichia coli. This plasmid was introduced into Erwinia chrysanthemi, and the pelC::mini-Mu insertion was substituted for the chromosomal allele by homologous recombination. This strain lacks the PLc isoenzyme. This Erwinia chrysanthemi strain has a Lac+ phenotype that is inducible by polygalacturonate, as are the wild-type PL activities.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andro T., Chambost J. P., Kotoujansky A., Cattaneo J., Bertheau Y., Barras F., Van Gijsegem F., Coleno A. Mutants of Erwinia chrysanthemi defective in secretion of pectinase and cellulase. J Bacteriol. 1984 Dec;160(3):1199–1203. doi: 10.1128/jb.160.3.1199-1203.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bertheau Y., Madgidi-Hervan E., Kotoujansky A., Nguyen-The C., Andro T., Coleno A. Detection of depolymerase isoenzymes after electrophoresis or electrofocusing, or in titration curves. Anal Biochem. 1984 Jun;139(2):383–389. doi: 10.1016/0003-2697(84)90022-8. [DOI] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casadaban M. J., Chou J. In vivo formation of gene fusions encoding hybrid beta-galactosidase proteins in one step with a transposable Mu-lac transducing phage. Proc Natl Acad Sci U S A. 1984 Jan;81(2):535–539. doi: 10.1073/pnas.81.2.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chatterjee A. K., Buchanan G. E., Behrens M. K., Starr M. P. Synthesis and excretion of polygalacturonic acid trans-eliminase in Erwinia, Yersinia, and Klebsiella species. Can J Microbiol. 1979 Jan;25(1):94–102. doi: 10.1139/m79-014. [DOI] [PubMed] [Google Scholar]
  7. Chatterjee A. K., Starr M. P. Donor strains of the soft-rot bacterium Erwinia chrysanthemi and conjugational transfer of the pectolytic capacity. J Bacteriol. 1977 Dec;132(3):862–869. doi: 10.1128/jb.132.3.862-869.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chatterjee A. K., Starr M. P. Genetics of Erwinia species. Annu Rev Microbiol. 1980;34:645–676. doi: 10.1146/annurev.mi.34.100180.003241. [DOI] [PubMed] [Google Scholar]
  9. Collmer A., Bateman D. F. Impaired induction and self-catabolite repression of extracellular pectate lyase in Erwinia chrysanthemi mutants deficient in oligogalacturonide lyase. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3920–3924. doi: 10.1073/pnas.78.6.3920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gijsegem F., Toussaint A., Schoonejans E. In vivo cloning of the pectate lyase and cellulase genes of Erwinia chrysanthemi. EMBO J. 1985 Mar;4(3):787–792. doi: 10.1002/j.1460-2075.1985.tb03698.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hofnung M., Jezierska A., Braun-Breton C. lamB mutations in E. coli K12: growth of lambda host range mutants and effect of nonsense suppressors. Mol Gen Genet. 1976 May 7;145(2):207–213. doi: 10.1007/BF00269595. [DOI] [PubMed] [Google Scholar]
  12. Hugouvieux-Cotte-Pattat N., Robert-Baudouy J. Lactose metabolism in Erwinia chrysanthemi. J Bacteriol. 1985 Apr;162(1):248–255. doi: 10.1128/jb.162.1.248-255.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kotoujansky A., Diolez A., Boccara M., Bertheau Y., Andro T., Coleno A. Molecular cloning of Erwinia chrysanthemi pectinase and cellulase structural genes. EMBO J. 1985 Mar;4(3):781–785. doi: 10.1002/j.1460-2075.1985.tb03697.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pugsley A. P., Schnaitman C. A. Factors affecting the electrophoretic mobility of the major outer membrane proteins of Escherichia coli in polyacrylamide gels. Biochim Biophys Acta. 1979 Nov 23;581(1):163–178. doi: 10.1016/0005-2795(79)90233-2. [DOI] [PubMed] [Google Scholar]
  16. Starr M. P., Chatterjee A. K. The genus Erwinia: enterobacteria pathogenic to plants and animals. Annu Rev Microbiol. 1972;26:389–426. doi: 10.1146/annurev.mi.26.100172.002133. [DOI] [PubMed] [Google Scholar]
  17. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]

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