Biopolym. Cell. 2000; 16(5):356-362.
Structure and Function of Biopolymers
Cloning and structural analysis of the Klebsiella oxytoca VN13 peh gene
1Kovtunovich G. V., 1Lar O. V., 1Kozyrovska N. O.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


The nucleotide sequence of the promoter part and 1450 bp of the structural part of the endophytic bacterium Klebsiella oxytoca VN13 peh gene which probably takes part in the penetration of bacteria into the plant root tissue has been determined. The levels of homology of both DNA and amino acid sequences of the protein coded and appropriate sequences of different bacteria have been analysed. Cloning and structure analyse of Klebsiella oxytoca VN13 peh gene


[1] Colmer A, Keen NT. The Role of Pectic Enzymes in Plant Pathogenesis. Annu Rev Phytopathol. 1986;24(1):383–409.
[2] Roberts DP, Denny TP, Schell MA. Cloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity. J Bacteriol. 1988;170(4):1445-51.
[3] Schell MA, Roberts DP, Denny TP. Analysis of the Pseudomonas solanacearum polygalacturonase encoded by pglA and its involvement in phytopathogenicity. J Bacteriol. 1988;170(10):4501-8.
[4] Huang JH, Schell MA. DNA sequence analysis of pglA and mechanism of export of its polygalacturonase product from Pseudomonas solanacearum. J Bacteriol. 1990;172(7):3879-87.
[5] Hinton JC, Gill DR, Lalo D, Plastow GS, Salmond GP. Sequence of the peh gene of Erwinia carotovora: homology between Erwinia and plant enzymes. Mol Microbiol. 1990;4(6):1029-36.
[6] Saarilahti HT, Heino P, Pakkanen R, Kalkkinen N, Palva I, Palva ET. Structural analysis of the pehA gene and characterization of its protein product, endopolygalacturonase, of Erwinia carotovora subspecies carotovora. Mol Microbiol. 1990;4(6):1037-44.
[7] Lei SP, Lin HC, Wang SS, Higaki P, Wilcox G. Characterization of the Erwinia carotovora peh gene and its product polygalacturonase. Gene. 1992;117(1):119-24.
[8] Liu Y, Chatterjee A, Chatterjee AK. Nucleotide sequence and expression of a novel pectate lyase gene (pel-3) and a closely linked endopolygalacturonase gene (peh-1) of Erwinia carotovora subsp. carotovora 71. Appl Environ Microbiol. 1994;60(7):2545-52.
[9] Brooks AD, He SY, Gold S, Keen NT, Collmer A, Hutcheson SW. Molecular cloning of the structural gene for exopolygalacturonate lyase from Erwinia chrysanthemi EC16 and characterization of the enzyme product. J Bacteriol. 1990;172(12):6950-8.
[10] Herlache TC, Hotchkiss AT Jr, Burr TJ, Collmer A. Characterization of the Agrobacterium vitis pehA gene and comparison of the encoded polygalacturonase with the homologous enzymes from Erwinia carotovora and Ralstonia solanacearum. Appl Environ Microbiol. 1997;63(1):338-46.
[11] Gonzalez CF, Pettit EA, Valadez VA, Provin EM. Mobilization, cloning, and sequence determination of a plasmid-encoded polygalacturonase from a phytopathogenic Burkholderia (Pseudomonas) cepacia. Mol Plant Microbe Interact. 1997;10(7):840-51.
[12] Flego D, Pirhonen M, Saarilahti H, Palva TK, Palva ET. Control of virulence gene expression by plant calcium in the phytopathogen Erwinia carotovora. Mol Microbiol. 1997;25(5):831-8.
[13] Kovtunovych GL, Lar OV, Kordyum VA, Kleiner D, Kozyrovska NO. Enhancing the internal plant colonization rate with endophytic nitrogen-fixing bacteria. Biopolym Cell. 1999;15(4):300-5.
[14] Pridmore RD. New and versatile cloning vectors with kanamycin-resistance marker. Gene. 1987;56(2-3):309-12.
[15] Miller JH. Experiments in molecular genetics. Cold Spring Harbor Laboratory, 1972, 466 p.
[16] 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.
[17] Nasuno S, Starr MP. Polygalacturonase of Erwinia carotovora. J Biol Chem. 1966;241(22):5298-306.
[18] Methods of studying the properties and cellulolytic enzymes. Ed. SD Varfolomeev. M.,(Itogi nauki i tekhniki; Biotekhnologiya; Vol 25) 1990: 138-140.
[19] Marmur J. A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol. 1961;3(2):208-18.
[20] Sambrook J, Fritsch EF, Maniatis T. Molecular cloning. A laboratory manual. The second edition. New York: Cold Spring Harbor Lab., 1989 p.
[21] 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.
[22] Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977;74(12):5463-7.
[23] Liao CH, Revear L, Hotchkiss A, Savary B. Genetic and biochemical characterization of an exopolygalacturonase and a pectate lyase from Yersinia enterocolitica. Can J Microbiol. 1999;45(5):396-403.
[24] He SY, Collmer A. Molecular cloning, nucleotide sequence, and marker exchange mutagenesis of the exo-poly-alpha-D-galacturonosidase-encoding pehX gene of Erwinia chrysanthemi EC16. J Bacteriol. 1990;172(9):4988-95.
[25] Huang Q, Allen C. An exo-poly-alpha-D-galacturonosidase, PehB, is required for wild-type virulence of Ralstonia solanacearum. J Bacteriol. 1997;179(23):7369-78.
[26] Chatterjee AK, Buchanan GE, Behrens MK, Starr MP. Synthesis and excretion of polygalacturonic acid trans-eliminase in Erwinia, Yersinia, and Klebsiella species. Can J Microbiol. 1979;25(1):94-102.
[27] Oliver D. Protein secretion in Escherichia coli. Annu Rev Microbiol. 1985;39:615-48.
[28] von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985;184(1):99-105.