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Construction of new yeast vectors and cloning of the nif (nitrogen fixation) gene cluster of Klebsiella pneumoniae in yeast

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Summary

Two vectors, termed pG63.11 (7.6 Kb) and pHCG3 (9.6 Kb), suitable for yeast transformation have been constructed. The pHCG3 vector has cosmid properties. Both vectors contain a single 3.3 Kb EcoRI-HindIII fragment of yeast origin which carries the yeast URA3 gene (1.1 Kb) and the origin of replication of the 2 µm plasmid (2.2 Kb). They confer ampicillin resistance and they contain 5 unique EcoRI,HpaI,HindIII,BamHI and SalI restriction sites. Cosmid pHCG3 was used to clone the nitrogen fixation (nif) gene cluster of Klebsiella pneumoniae carried by twoHindIII fragments of 17 and 26 Kb, respectively. The resulting cosmid, termed pGPC875 (53 Kb) which conferred a Nif+ phenotype to Escherichia coli, was introduced in yeast by transformation. No acetylene reduction activity was detectable in the transformants. However it was shown that the entire information for nitrogen fixation can be replicated and maintained intact in yeast for more than 50 generations of growth.

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References

  • Aigle M, Chevallier MR (1979) Febs Lett 108:179–180

    Google Scholar 

  • Apelbaum ER, Kramer RA (1980) Mol Gen Genet 179:349–134

    Google Scholar 

  • Bach ML, Lacroute F, Botstein D (1979) Proc Natl Acad Sci USA 76:386–390

    Google Scholar 

  • Beggs J (1978) Nature 275:104–108

    Google Scholar 

  • Blanc H, Gerbaud C, Slonimski P, Guérineau M (1979) Mol Gen Genet 176:335–342

    Google Scholar 

  • Birboim HC, Doly J (1979) Nucl Acids Res 7:1513–1523

    Google Scholar 

  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Gene 2:95–113

    Google Scholar 

  • Broach JR, Hicks JB (1980) Cell 21:501–508

    Google Scholar 

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

    Google Scholar 

  • Cannon FC, Dixon RA, Postgate JR, Primrose SB (1974) J Gen Microbiol 80:227–239

    Google Scholar 

  • Cannon FC, Riedel GE, Ausubel FM (1979) Mol Gen Genet 172:59–66

    Google Scholar 

  • Chevallier MR, Bloch JC, Lacroute F (1980) Gene 11:11–19

    Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Proc Natl Acad Sci USA 69:2110–2114

    Google Scholar 

  • Colbere-Garapin F, Chousterman S, Horodiceanu F, Kourilski P, Garapin AC (1979) Proc Natl Acad Sci USA 76:3755–3759

    Google Scholar 

  • Collins, J, Brüning HJ (1978) Gene 4:85–107

    Google Scholar 

  • Collins J, Hohn B (1978) Proc Natl Acad Sci USA 75:4242–4246

    Google Scholar 

  • Dixon RA, Cannon F, Kondorosi A (1976) Nature 260:268–271

    Google Scholar 

  • Dixon R, Eady RR, Espin G, Hill S, Iaccarino M, Kahn D, Merrick M (1980) Nature 286:128–132

    Google Scholar 

  • Dixon RA, Kennedy C, Kondorosi A, Krishnapillai V, Merrick M (1977) Mol Gen Genet 157:189–198

    Google Scholar 

  • Elmerich C, Houmard J, Sibold L, Manheimer I, Charpin N (1978) Mol Gen Genet 165:181–189

    Google Scholar 

  • Gerbaud C, Fournier P, Blanc H, Aigle M, Heslot H, Guerineau M (1979) Gene 5:233–253

    Google Scholar 

  • Guérineau M, Grandchamp C, Slonimski PP (1976) Proc Natl Acad Sci USA 73:3030–3034

    Google Scholar 

  • Guérineau M, Slonimski PP, Avner PR (1974) Biochem Biophys Res Comm 61:462–469

    Google Scholar 

  • Guerry YP, Le Blanc DJ, Falkow S (1973) J Bacteriol 116:1064–1066

    Google Scholar 

  • Hanahan D, Meselson M (1980) Gene 6:63–67

    Google Scholar 

  • Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933

    Google Scholar 

  • Hohn B, Collins J (1980) Gene 10:291–298

    Google Scholar 

  • Hohn B, Hinnen A (1980) Cloning with cosmids in E. coli and yeast. In: Setlow JK, Hollaender A (eds) Genetic Engineering, 2. Plenum Press, New York, p 169

    Google Scholar 

  • Hsiao CL, Carbon J (1979) Proc Natl Acad Sci USA 76:3829–3833

    Google Scholar 

  • Humphreys GO, Willshaw GA, Anderson ES (1975) Biochim Biophys Acta 383:457–463

    Google Scholar 

  • Leonardo JM, Goldberg RB (1980) J Bacteriol 142:99–110

    Google Scholar 

  • Losson R, Lacroute F (1979) Proc Natl Acad Sci USA 76:5134–5137

    Google Scholar 

  • MacNeil D, Brill WJ (1980) J Bacteriol 144:744–757

    Google Scholar 

  • MacNeil T, MacNeil D, Roberts GP, Supiano MA, Bril WJ (1978) J Bacteriol 136:821–829

    Google Scholar 

  • Merrick M, Filser M, Dixon R, Elmerich C, Sibold L, Houmard J (1980) J Gen Microbiol 117:509–520

    Google Scholar 

  • Miller JH (1972) In: Experiments in molecular genetics, Cold Spring Harbor Laboratory (ed), Cold Spring Harbor

    Google Scholar 

  • Murray NE, Brammar WJ, Murray K (1977) Mol Gen Genet 150:53–61

    Google Scholar 

  • Panthier JJ, Fournier P, Heslot, H, Rambach A (1980) Curr Gen 2:109–113

    Google Scholar 

  • Petes TD (1980) Ann Rev Biochem 49:445–476

    Google Scholar 

  • Petes TD, Broach JR, Wensink PC, Fink GR, Botstein D (1978) Gene 4:37–49

    Google Scholar 

  • Philipsen P, Kramer RA, Davis RW (1978) J Mol Biol 123:371–386

    Google Scholar 

  • Pühler A, Burkardt HJ, Klipp W (1979) Mol Gen Genet 176:11–24

    Google Scholar 

  • Riedel GE, Ausubel FM, Cannon FC (1979) Proc Natl Acad Sci USA 76:2866–2870

    Google Scholar 

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

    Google Scholar 

  • Sanger F, Coulson AR, Friedman T, Air GM, Barrel BG, Brown NL, Fiddes JC, Hutchison III CA, Slocombe PM, Smith M (1978) J Mol Biol 125:225–246

    Google Scholar 

  • Shanmugam KT, Chan I, Morandi C (1975) Biochim Biophys Acta 408:101–111

    Google Scholar 

  • Sibold L, Melck D, Elmerich C (1981) FEMS Microbiol Lett 10:37–41

    Google Scholar 

  • Streicher SL, Shanmugam KT, Ausubel F, Morandi C, Goldberg RB (1974) J Bacteriol 120:815–821

    Google Scholar 

  • Storms RK, McNeil JB, Kandelar PS, Parker AJ, Friesen JD (1979) J Bacteriol 140:73–82

    Google Scholar 

  • Struhl K, Stinchomb DT, Scherrer S, Davis RW (1979) Proc Natl Acad Sci USA 75:1035–1039

    Google Scholar 

  • Vogt VM (1973) Eur J Biochem 33:192–200

    Google Scholar 

Download references

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Author notes

  1. Nicole Tandeau de Marsac

    Present address: Unité de Physiologie Microbienne, Institut Pasteur, France

Authors and Affiliations

  1. Laboratoire de Biologie Génétique et Moléculaire, Université de Paris Sud, Bâtiment 400, 91405, Orsay, France

    Claude Gerbaud & Michel Guérineau

  2. Unité de Physiologic Cellulaire, Département de Biochimie et Génétique Moléculaire, Institut Pasteur, 28 rue du Dr. Roux, 75724, Paris Cedex 15, France

    Claudine Elmerich, Nicole Tandeau de Marsac, Patrick Chocat, Nicole Charpin & Jean-Paul Aubert

Authors
  1. Claude Gerbaud
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  2. Claudine Elmerich
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  3. Nicole Tandeau de Marsac
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  4. Patrick Chocat
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  5. Nicole Charpin
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  6. Michel Guérineau
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  7. Jean-Paul Aubert
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Gerbaud, C., Elmerich, C., Tandeau de Marsac, N. et al. Construction of new yeast vectors and cloning of the nif (nitrogen fixation) gene cluster of Klebsiella pneumoniae in yeast. Curr Genet 3, 173–180 (1981). https://doi.org/10.1007/BF00429819

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  • DOI: https://doi.org/10.1007/BF00429819

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