Expression of invertase activity in Yarrowia lipolytica and its use as a selective marker
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Summary
Few selective markers are available for the transformation of the industrial yeast Yarrowia lipolytica, and those that are require the use of specialized hosts (e.g., auxotrophs, antibiotic sensitive). To enable the transformation of any Y. lipolytica strain, we used the property that Y. lipolytica cannot use sucrose as a sole carbon source. We have constructed a gene fusion where the Saccharomyces cerevisiae SUC2 gene is placed under the control of the promoter and signal sequence of the Y. lipolytica XPR2 gene, which encodes an Alkaline Extracellular Protease (AEP). Strains bearing this fusion express invertase activity and grow on sucrose as a carbon source. The activity follows the same regulation as does the alkaline extracellular protease, is secreted into the periplasm and confers a Suc+ phenotype. It was shown that this chimeric gene could be used as a dominant marker for transformation in a one-step procedure.
Key words
Yarrowia lipolytica Invertase Secretion TransformationPreview
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References
- Ahearn DG, Meyers SP, Nichols RA (1968) Appl Microbiol 16:1370–1374Google Scholar
- Basset J, Mortimer R (1973) J Bacteriol 114:894–896Google Scholar
- Boyer HW, Roulland-Dussoix D (1969) J Mol Biol 41:459–472Google Scholar
- Carlson M, Botstein D (1982) Cell 28:145–154Google Scholar
- Cohen JD, Eccleshall TR, Needlemann RB, Federoff H, Buchferer BA, Marmur J (1980) Proc Natl Acad Sci USA 77:1078–1082Google Scholar
- Dagert M, Ehrlich SD (1979) Gene 6:23–28Google Scholar
- Davidow LS, Apostolakos D, O'Donnell MM, Proctor AR (1985) Curr Genet 10:39–48Google Scholar
- Davidow LA, Franke AE, DeZeeuw JR (1987) European Patent Application 220864Google Scholar
- Davidow LS, O'Donnell MM, Kaczmarek FS, Pereira DA, DeZeeuw JR, Franke AE (1987) J Bacteriol 169:4621–4629Google Scholar
- Efimov VA, Burykova AA, Reverdato SV, Chakhmakhcheva OG, Ovchinnikov YA (1983) Nucleic Acids Res 11:8369Google Scholar
- Gaillardin C, Ribet AM (1987) Curr Genet 11:369–375Google Scholar
- Gaillardin CM, Charoy V, Heslot H (1973) Arch Microbiol 92:69–83Google Scholar
- Gaillardin C, Ribet AM, Heslot H (1985) Curr Genet 10:49–58Google Scholar
- Hecht SM (1986) Fed Proc 45:2784–2791Google Scholar
- Holmes DS, Quigley M (1981) Anal Biochem 114:193Google Scholar
- Jimenez A, Davies J (1980) Nature 287:869–871Google Scholar
- Kamada M, Ogura S, Oda K, Murao S (1972) Agric Biol Chem 36:171–175Google Scholar
- Klug MJ, Markovetz AJ (1967) J Bacteriol 93:1847–1851Google Scholar
- Klug MJ, Markovetz AJ (1969) Biotechnol Bioeng 11:427–440Google Scholar
- Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
- Matoba S, Fukayama J, Wing R, Ogrydziak DM (1988) Mol Cell Biol 8:4904–4916Google Scholar
- Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
- Ogrydziak DM, Scharf SJ (1982) J Gen Microbiol 128:1225–1234Google Scholar
- Ogrydziak DM, Demain AL, Tannenbaum SR (1977) Biochem Biophys Acta 497:525–538Google Scholar
- Robinson JS, Klionsky DJ, Banta LM, EMR SD (1988) Mol Cell Biol 8:4936–4948Google Scholar
- Sanger F, Nicklen S, Couloon AR (1977) Proc Natl Acad Sci USA 74:5463–5467Google Scholar
- Sherman F, Fink GR, Hicks JB (1979) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 90–92Google Scholar
- Taussig R, Carlson M (1983) Nucleic Acids Res 11:1943–1954Google Scholar
- Taylor JW, Ott J, Eckstein F (1985) Nucleic Acids Res 13:8764–8785Google Scholar
- Tobe ST, Takami S, Ikeda S, Mitsugi K (1976) Agric Biol Chem 40:1037–1092Google Scholar
- von Heijne G (1986) Nucleic Acids Res 14:4683–4690Google Scholar
- Werner W, Rey HG, Wielinger H (1970) Z Anal Chem 252:224–225Google Scholar
- Xuan J-W, Fournier P, Gaillardin C (1988) Curr Genet 14:15–21Google Scholar
- Yanisch-Perron C, Vigra J, Messing J (1985) Gene 33:103–119Google Scholar