Biopolym. Cell. 2005; 21(1):55-59.
http://dx.doi.org/10.7124/bc.0006DC
Molecular and Cell Biotechnologies
Role of the pelX gene of Klebsiella oxytoca VN13 in pectinolysis
1Lar O. V., 1Kovtunovych G. L., 1Kozyrovska N. O.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

An insertionally inactivated copy of the pelX gene of K. oxytoca VN13 was used for the wild type gene substitution by reciprocal recombination. The presence in K. oxytoca VN13 genome of the additional pectate lyase gene with nucleotide sequence differing from the pelX gene was revealed. A considerably lower level of the pectate lyase activity of the mutant K. oxytoca VN13 strain compared to the wild type strain was determined. It was shown, that the pelX gene expression is not essential for polygalacturonate assimilation by this bacterium.
Keywords: Klebsiella oxytoca, pectinolysis, insersion mu­ tagenesis, the pelX gene
Full text: (PDF, in Ukrainian)

References

[1] K Kovtunovych G, Lar O, Kamalova S, Kordyum V, Kleiner D, Kozyrovska N. Correlation between pectate lyase activity and ability of diazotrophic Klebsiella oxytoca VN13 to penetrate into plant tissues. Plant Soil. 1999;215(1):1–6.
[2] Kovtunovich GV, Lar OV, Kozyrovska NO. Cloning and structural analysis of the Klebsiella oxytoca VN13 peh gene. Biopolym Cell. 2000; 16(5):356-62.
[3] Lar HV, Kovtunovich GL, Kozyrovska NO. Cloning and analysis of the pectate lyase pelX gene of Klebsiella oxytoca VN13. Biopolym Cell. 2002; 18(5):417-22.
[4] Kovtunovych GL, Lar OV, Kozyrovska NO. Role of the exopolygalacturonase gene in interaction of Klebsiella oxytoca VN13 with wheat roots. Biopolym Cell. 2002; 18(4):319-23.
[5] Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103-19. ttp://
[6] Benes V, Hostomsk? Z, Arnold L, Paces V. M13 and pUC vectors with new unique restriction sites for cloning. Gene. 1993;130(1):151-2.
[7] Fellay R, Frey J, Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147-54.
[8] Penfold RJ, Pemberton JM. An improved suicide vector for construction of chromosomal insertion mutations in bacteria. Gene. 1992;118(1):145-6.
[9] Miller JH. Experiments in molecular genetics. Cold Spring Harbor Laboratory, 1972, 466 p.
[10] Starr MP, Chatterjee AK, Starr PB, Buchanan GE. Enzymatic degradation of polygalacturonic acid by Yersinia and Klebsiella species in relation to clinical laboratory procedures. J Clin Microbiol. 1977;6(4):379-86.
[11] Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: a laboratory manual. New York: Cold Spring Harbor Lab., 1989.
[12] Nishimura A, Morita M, Nishimura Y, Sugino Y. A rapid and highly efficient method for preparation of competent Escherichia coli cells. Nucleic Acids Res. 1990;18(20):6169.
[13] Walker MJ, Pemberton JM. Construction of a transposon containing a gene for polygalacturonate trans-eliminase from Klebsiella oxytoca. Arch Microbiol. 1987;146(4):390-5.
[14] Hugouvieux-Cotte-Pattat N, Condemine G, Nasser W, Reverchon S. Regulation of pectinolysis in Erwinia chrysanthemi. Annu Rev Microbiol. 1996;50:213-57.