Current Genetics

, Volume 24, Issue 6, pp 491–495 | Cite as

Cloning and manipulation of the Schizosaccharomyces pombe his7+gene as a new selectable marker for molecular genetic studies

  • Ethel Apolinario
  • Mary Nocero
  • Mei Jin
  • Charles S. Hoffman
Original Articles


We have cloned the his7+ gene of the fission yeast Schizosaccharomyces pombe by complementation of the recessive mutant allele his7-366. The his7+ gene is able to complement a mutation of the Escherichia coli hisI gene, suggesting that his7+ encodes a phosphoribosyl-AMP cyclohydrase. Subcloning experiments localize the gene to a 1.9-kb XbaI-BglII fragment. We describe the construction of plasmids to facilitate the use of his7+ as a selectable marker in S. pombe studies. Plasmid pEA2 carries his7+ cloned into the pUC18 polylinker. From either pEA2 or the original his7+ clone, pMN1, fragments carrying his7+ can be isolated using a variety of restriction enzymes for the construction of gene disruptions. Plasmid pEA500 is a cloning vector that carries his7+ and ars1, yet retains the ability to use the blue/white color screen to identify recombinants.

Key words

Fission yeast Selectable marker Gene disruptions Cloning vector 


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  1. Ausubel, F, Brent R, Kingston R, Moore D, Smith J, Seidman J, Struhl K (eds) (1987) Current protocols in molecular biology. Wiley, New YorkGoogle Scholar
  2. Bach ML (1987) Curr Genet 12:527–534Google Scholar
  3. Bach ML, Lacroute F, Botstein D (1979) Proc Natl Acad Sci USA 76:386–390Google Scholar
  4. Beach D, Nurse P (1981) Nature 290:140–142Google Scholar
  5. Beach D, Rogers L, Gould J (1985) Curr Genet 10:297–311Google Scholar
  6. Boeke JD, Lacroute F, Fink GR (1984) Mol Gen Genet 197:345–346Google Scholar
  7. Boeke JD, Trueheart J, Natsoulis G, Fink GR (1987) Methods Enzymol 154:164–175Google Scholar
  8. Botstein D, Falco SC, Stewart S, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis R (1979) Gene 8:17–24Google Scholar
  9. Boyer HW, Roulland-Dussiox, D (1969) J Mol Biol 41:458–472Google Scholar
  10. Broach JR, Strathern JN, Hicks JB (1979) Gene 8:121–133Google Scholar
  11. Casadaban MJ, Cohen SN (1980) J Mol Biol 138:179–207Google Scholar
  12. Chiariotti L, Alifano P, Carlomagno MS, Bruni CB (1986) Mol Gen Genet 203:382–388Google Scholar
  13. Cottarel G, Beach D, Deuschle U (1993) Curr Genet 23:547–578Google Scholar
  14. Elble R (1992) BioTechniques 13:18–20Google Scholar
  15. Goldschmidt EP, Cater MS, Matney TS, Butler MA, Greene A (1970) Genetics 66:219–229Google Scholar
  16. Grimm C, Kohli J, Murray J, Maundrell K (1988) Mol Gen Genet 215:81–86Google Scholar
  17. Gutz H, Heslot H, Leupold U, Loprieno N (1974) In: King RC (ed) Handbook of genetics. Plenum Press, New York, pp 395–446Google Scholar
  18. Hoffman CS, Winston F (1987) Gene 57:267–272Google Scholar
  19. Hoffman CS, Winston F (1990) Genetics 124:807–816Google Scholar
  20. Hoffman CS, Winston F (1991) Genes Dev 5:561–571Google Scholar
  21. Kikuchi Y, Kitazawa Y, Shimatake H, Yamamoto Y (1988) Curr Genet 14:375–379Google Scholar
  22. Kohli J (1987) Curr Genet 11:575–589Google Scholar
  23. Losson R, Lacroute F (1983) Cell 32:371–377Google Scholar
  24. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  25. Maundrell K, Wright APH, Piper M, Shall S (1985) Nucleic Acids Res 13:3711–3722Google Scholar
  26. Norrander J, Kempe T, Messing J (1983) Gene 26:101–106Google Scholar
  27. Ratzkin B, Carbon J (1977) Proc Natl Acad Sci USA 74:487–491Google Scholar
  28. Rothstein RJ (1983) Methods Enzymol 101:202–211Google Scholar
  29. Russell P, Nurse P (1987) Cell 49:559–567Google Scholar
  30. Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  31. Southern EM (1975) J Mol Biol 98:503–517Google Scholar
  32. Wright A, Maundrell K, Heyer HD, Beach D, Nurse P (1986) Plasmid 15:156–158Google Scholar
  33. Yamawaki-Kataoka Y, Tamaoki T, Choe HR, Tanaka H, Kataoka T (1989) Proc Natl Acad Sci USA 86:5693–5697Google Scholar
  34. Young D, Riggs M, Fields J, Vojtek A, Broek D, Wigler M (1989) Proc Natl Acad Sci USA 86:7989–7993Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Ethel Apolinario
    • 1
  • Mary Nocero
    • 1
  • Mei Jin
    • 1
  • Charles S. Hoffman
    • 1
  1. 1.Department of BiologyBoston CollegeChestnut HillUSA

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