Current Microbiology

, Volume 23, Issue 3, pp 123–129 | Cite as

Cloning and expression inEscherichia coli of arecA-like gene fromZymomonas mobilis

  • T. Karunakaran
  • P. Gunasekaran


Intergeneric complementation ofEscherichia coli recA mutants was used to identify recombinant plasmids, within a genomic library derived fromZymomonas mobilis, that carryZ. mobilis recA-like gene. Screening of 1100 individualE. coli strains revealed four clones expressing therecA+ character. On restriction analysis, all four recombinant plasmids were found to be related and to exhibit a common 6.7-kb fragment. Consequently, one of the four recombinant plasmids, pZR27, was selected for further characterization. When introduced intoE. coli recA mutants, pZR27 restored resistance to methyl methane sulfonate, mitomycin-C, and UV irradiation, as well as recombination proficiency when measured by standard Hfr-mediated conjugation. The clonedrecA-like gene also restored the spontaneous and mitomycin-C-induced phage production. The origin of the insert in pZR27 from the chromosome ofZ. mobilis was confirmed by Southern transfer and DNA hybridization. However, no homology was found between therecA ofE. coli andZ. mobilis chromosomal insert DNA. TheZ. mobilis recA-like gene also encoded a major polypeptide of 38-kDa on SDS-PAGE.


Recombination Recombinant Plasmid Restriction Analysis Genomic Library Methane Sulfonate 
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Literature Cited

  1. 1.
    Better M, Helinski DR (1984) Isolation and characterization of therecA gene ofRhizobium meliloti. J Bacteriol 155:311–316Google Scholar
  2. 2.
    Boyer HW, Roueland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA inEscherichia coli. J Mol Biol 41:459–472PubMedGoogle Scholar
  3. 3.
    Brestic-Goachet N, Gunasekaran P, Cami B, Baratti J (1990) Transfer and expression of aBacillus licheniformis α-amylase gene inZymomonas mobilis. Arch Microbiol 153:219–225Google Scholar
  4. 4.
    Cohen G (1983) Electron microscopy study of early lytic replication forms of bacteriophage P1 DNA. Virology 131:159–170PubMedGoogle Scholar
  5. 5.
    Craig NL, Roberts JW (1980)E. coli recA protein-directed cleavage of phage repressor requires polynucleotide. Nature (London) 283:26–30PubMedGoogle Scholar
  6. 6.
    Geoghegan CM, Houghton JA (1987) Molecular cloning and isolation of a cyanobacterial gene which increases the UV and methyl methane sulfonate survival ofrecA strains ofEscherichia coli K12. J Gen Microbiol 133:119–126PubMedGoogle Scholar
  7. 7.
    Goldberg I, Mekalanos JJ (1986) Cloning of theVibrio cholerae recA gene and construction of aVibrio cholerae recA mutant. J Bacteriol 165:715–722PubMedGoogle Scholar
  8. 8.
    Goodman HJK, Parker JR, Southern JA, Woods DR (1987) Cloning and expression inEscherichia coli of arecA like gene fromBacteroides fragilis. Gene 58:265–271PubMedGoogle Scholar
  9. 9.
    Gunasekaran P, Karunakaran T, Cami B, Mukundan AG, Presiozi L, Baratti J (1990) Cloning and sequencing ofsacA gene: characterization of a sucrase fromZymomonas mobilis. J Bacteriol 172:6727–6735PubMedGoogle Scholar
  10. 10.
    Hanawalt PC, Cooper PK, Ganesan AK, Smith CA (1979) DNA repair in bacteria and mammalian cells. Annu Rev Biochem 48:783–786PubMedGoogle Scholar
  11. 11.
    Koomey JM, Falkow S (1987) Cloning of therecA gene ofNeisseria gonorrhoeae and construction of gonococcalrecA mutants. J Bacteriol 169:790–795PubMedGoogle Scholar
  12. 12.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685PubMedGoogle Scholar
  13. 13.
    Lejeune A, Eveleigh DE, Colson C (1988) Expression of an endoglucanase gene ofPseudomonas fluorescens var.cellulosa inZymomonas mobilis. FEMS Microbiol Lett 49:363–366Google Scholar
  14. 14.
    Little JW (1984) Autodigestion oflexA and phage repressors. Proc Natl Acad Sci USA 81:1375–1379PubMedGoogle Scholar
  15. 15.
    Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory.Google Scholar
  16. 16.
    Maynard MT, Kuramitsu HK (1979) Purification and antigenic properties of intracellular invertase fromStreptococcus mutans. Infect Immun 23:873–883PubMedGoogle Scholar
  17. 17.
    Miller JH (1972) Experiments in molecular genetics. New York: Oxford University PressGoogle Scholar
  18. 18.
    Owttrim GW, Coleman JR (1987) Molecular cloning of arecA like gene from the cyanobacteriumAnabaena variabilis. J. Bacteriol. 169:1824–1829PubMedGoogle Scholar
  19. 19.
    Rogers PL, Lee KJ, Skotnicki ML, Tribe DE (1982) Ethanol production byZymomonas mobilis. Adv Biochem Eng 23:37–84Google Scholar
  20. 20.
    Ruther U (1982) pUR250 allows rapid chemical sequencing of both DNA strands of its inserts. Nucleic Acids Res 10:5765–5772PubMedGoogle Scholar
  21. 21.
    Shibata T, Dasgupta C, Cunningham RP, Williams JGK, Osber L, Radding CM (1981) Homologous pairing in genetic recombination. The pairing reaction catalyzed byEscherichia coli recA protein. J Biol Chem 256:7565–7572PubMedGoogle Scholar
  22. 22.
    Walker GC (1984) Mutagenesis and inducible responses to deoxyribonucleic acid damage inEscherichia coli. Microbiol Rev 48:60–93PubMedGoogle Scholar
  23. 23.
    Walker GC (1985) Inducible DNA repair systems. Ann Rev Biochem 54:425–457PubMedGoogle Scholar
  24. 24.
    West SS, Cassuto E, Howard-Flanders P (1981)RecA protein promotes homologous pairing and strand exchange reactions between duplex DNA molecules. Proc Natl Acad Sci USA 78:2100–2104PubMedGoogle Scholar
  25. 25.
    Yoon KH, Park SH, Pack MY (1988) Transfer ofBacillus subtilis endoβ-1,4-glucanase gene intoZymomonas anaerobia. Biotechnol Lett 10:213–216Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1991

Authors and Affiliations

  • T. Karunakaran
    • 1
  • P. Gunasekaran
    • 1
  1. 1.Department of Microbiology and Microbial Technology, School of Biological SciencesMadurai Kamaraj UniversityMaduraiIndia

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