Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Molecular cloning of the ARG7 gene of Schizosaccharomyces pombe encoding argininosuccinate lyase

  • 23 Accesses

  • 2 Citations

Summary

A gene bank of Sau3A partially restricted Schizosaccharomyces pombe DNA in YEp13 was used to transform an arg4 mutant of Saccharomyces cerevisiae. One colony was recovered which contained the YEp13 plasmid bearing a large insert complementing the argininosuccinate lyase (ASL) mutation. As shown by restriction mapping and subcloning experiments, the DNA sequence required for complementation is localized on a 2 kb BamHI-BamHI fragment. The plasmid complemented several S. cerevisiae arg4 mutants of independent origin and a S. pombe arg7 mutant lacking ASL. Low but significant ASL activities were detected in crude extracts of these transformants. No complementation of the E. coli argH mutant was observed. Southern blot hybridizations showed that the insert originates from the S. pombe genome. No cross-hybridization was found between this sequence and S. cerevisiae DNA. It can be concluded that the cloned DNA fragment bears the S. pombe ARG7 gene coding for ASL.

This is a preview of subscription content, log in to check access.

Abbreviations

ASL :

argininosuccinate lyase

bp :

base pair

kb :

kilobase pair

References

  1. Aves SJ, Durkacz BW, Carr A, Nurse P (1985) EMBO J 4:457–463

  2. Bach ML (1987) Curr Genet 12:527–534

  3. Barker DG (1987) Eur J Biochem 162:659–667

  4. Beach D, Nurse P (1981) Nature 290:140–142

  5. Beach D, Piper M, Nurse P (1982) Mol Gen Genet 187:326–329

  6. Beacham IR, Schweitzer BW, Warrick HM, Carbon J (1984) Gene 29:271–279

  7. Berse B, Dmochowska A, Skrzypek M, Weglenski P, Bates MA, Weiss RL (1983) Gene 25:109–117

  8. Boy-Marcotte E, Jacquet M (1982) Gene 20:433–440

  9. Bradford MM (1976) Anal Biochem 72:248–254

  10. Broach JR, Strathern JN, Hicks JB (1979) Gene 8:121–133

  11. Clarke L, Carbon J (1978) J Mol Biol 120:517–532

  12. Egel R, Kohli J, Thuriaux P, Wolf K (1980) Annu Rev Genet 14:77–108

  13. Henikoff S, Tatchell K, Hall BD, Nasmyth KA (1981) Nature 289:33–37

  14. Henry SA, Klig LS, Loewy BS (1984) Annu Rev Genet 18:207–231

  15. Hudock GA (1963) Biochem Biophys Res Commun 10:133–138

  16. Kaüfer NF, Simanis V, Nurse P (1985) Nature 318:78–80

  17. Kuranda MJ, Robbins PW (1987) Proc Natl Acad Sci USA 84:2585–2589

  18. Loppes R, Denis C (1983) Curr Genet 7:473–480

  19. Lorincz A (1984) Focus 6

  20. Losson R, Lacroute F (1983) Cell 32:371–377

  21. Maniatis T, Fritsch EF, Sambrook J (1982). In: Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratories, Cold Spring Harbor, NY

  22. Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) J Mol Biol 113:237–251

  23. Schimoda C, Uchira M, Kishida M, Fujioka H, Iino Y, Watanabe Y, Yamamoto M (1987) J Bacteriol 169:93–96

  24. Southern EM (1975) J Mol Biol 98:503–517

  25. Tschumper G, Carbon J (1982) J Mol Biol 156:293–307

Download references

Author information

Correspondence to Roland Loppes.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Remacle, J.E., Breyer, D. & Loppes, R. Molecular cloning of the ARG7 gene of Schizosaccharomyces pombe encoding argininosuccinate lyase. Curr Genet 14, 381–385 (1988). https://doi.org/10.1007/BF00419996

Download citation

Key words

  • Schizosaccharomyces pombe
  • Saccharomyces cerevisiae
  • Argininosuccinate lyase
  • Gene cloning