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Current Genetics

, Volume 20, Issue 3, pp 219–224 | Cite as

Cloning, sequencing and expression of the Schwanniomyces occidentalis NADP-dependent glutamate dehydrogenase gene

  • P. A. De Zoysa
  • I. F. Connerton
  • D. C. Watson
  • J. R. Johnston
Original Articles

Summary

The cloned NADP-specific glutamate dehydrogenase (GDH) genes of Aspergillus nidulans (gdhA) and Neurospora crassa (am) have been shown to hybridize under reduced stringency conditions to genomic sequences of the yeast Schwanniomyces occidentalis. Using 5′ and 3′ gene-specific probes, a unique 5.1 kb BclI restriction fragment that encompasses the entire Schwanniomyces sequence has been identified. A recombinant clone bearing the unique BclI fragments has been isolated from a pool of enriched clones in the yeast/E. coli shuttle vector pWH5 by colony hybridization. The idenity of the plasmid clone was confirmed by functional complementation of the Saccharomyces cerevisiae ghd-1 mutation. The nucleotide sequence of the Schw. occidentalis GDH gene, which consists of 1380 nucleotides in a continuous reading frame of 459 amino acids, has been determined. The predicted amino acid sequence shows considerable homology with GDH proteins from other fungi and significant homology with all other available GDH sequences.

Key words

Schwanniomyces occidentalis Expression in Saccharomyces cerevisiae NADP-Glutamate dehydrogenase DNA sequence 

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References

  1. Bennetzen JL, Hall BD (1982) J Biol Chem 257:3026–3031Google Scholar
  2. Dobson MJ, Tuite MF, Roberts NA, Kingsman AJ, Kingsman SM, Perkins RE, Conroy SC, Dunbar B, Fothergill LA (1982) Nucleic Acids Res 10:2625–2637Google Scholar
  3. Dohmen RJ, Strasser AWM, Zitomer RS, Hollenberg CP (1989) Curr Genet 15:319–325Google Scholar
  4. Dohmen RJ, Strasser AWM, Dahlems UM, Hollenberg CP (1990) Gene 95:111–121Google Scholar
  5. Feinberg AP, Vogelstein B (1983) Anal Biochem 132:6–13Google Scholar
  6. Ferguson J, Ho J-Y, Peterson TA, Reed SI (1986) Nucleic Acids Res 14:6681–6697Google Scholar
  7. Finlayson SD, Fleming C, Berry DR, Johnston JR (1991) Biotechnol Tech 5:13–18Google Scholar
  8. Grenson M, Dubois E, Piotrowska M (1974) Mol Gen Genet 128:73–85Google Scholar
  9. Gurr SJ, Hawkins AR, Drainas C, Kinghorny JR (1986) Curr Genet 10:761–766Google Scholar
  10. Hanahan D (1985) In: DNA cloning, a practical approach I. 109–136Google Scholar
  11. Hawkins AR, Gurr SJ, Montague P, Kinghorn JR (1989) Mol Gen Genet 218:105–111Google Scholar
  12. Ito H, Fukuda Y, Murata K, Kimura A (1983) J Bacteriol 153:163–168Google Scholar
  13. Johnston JR, Contopoulou CR, Mortimer RK (1988) Yeast 4:191–198Google Scholar
  14. Kinnaird JH, Fincham JRS (1983) Gene 26:253–260Google Scholar
  15. Kinnaird JH, Keighren MA, Kinsey JA, Eaton M, Fincham JRS (1982) Gene 20:387–396Google Scholar
  16. Klein RD, Favreau MA (1988) J Bacteriol 170 (12):5572–5578Google Scholar
  17. Klein RD, Roof LL (1988) Curr Genet 13:29–35Google Scholar
  18. Klein RD, Poorman RA, Favreau MA, Shea MH, Hatzbuhler NT, Nulf SC (1989) Curr Genet 16:145–152Google Scholar
  19. Kozak M (1981) Nucleic Acids Res 9:5233–5252Google Scholar
  20. McKenzie R, Schuchert P, Kilbey B (1987) Curr Genet 12:591–597Google Scholar
  21. Maniatis T, Fritsh EF, Sambrook J (1982) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  22. Moye WS, Amuro N, Rao JKM, Zalkin H (1985) J Biol Chem 260 (14):8502–8508Google Scholar
  23. Nagasu T, Hall BD (1985) Gene 37:247–253Google Scholar
  24. Prakash K, Seligy VL (1988) Gene 73:131–140Google Scholar
  25. Rashied I, Jornvall H, Stund H (1974) Eur J Biochem 41:603–606Google Scholar
  26. Rodriguez RL., Tait RC (1983) In: Recombinant DNA techniques: an introduction. Addison-Wesley Publishing Company, Reading, Mass., pp 168–169Google Scholar
  27. Russell PR, Hall BD (1983) J Biol Chem 258:143–149Google Scholar
  28. Roon RJ, Even HL, Larimore F (1974) J Bacterol 118:89–95Google Scholar
  29. Sambrook J, Fritsh EF, Maniatis T (1988) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  30. Sherman F, Fink GR, Hicks JB (1983) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  31. Strasser AWM, Selk R, Dohmen RJ, Niermann T, Bielfield M, Seeboth P, Tu G, Hollenberg CP (1989) Eur J Biochem 184:699–706Google Scholar
  32. Wingender E (1988) Nucleic Acids Res 16(3):1879–1902Google Scholar
  33. Wright A, Maundrell K, Heyer W-D, Beach D, Nurse P (1986) Plasmid 15:156–158Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • P. A. De Zoysa
    • 1
    • 2
  • I. F. Connerton
    • 1
  • D. C. Watson
    • 3
  • J. R. Johnston
    • 2
  1. 1.Department of MicrobiologyUniversity of ReadingReadingEngland, UK
  2. 2.Department of Bioscience and BiotechnologyUniversity of StrathclydeGlasgowScotland, UK
  3. 3.Chivas BrothersKeithScotland, UK

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