Summary
We present here the first report of a transformation system developed for the filamentous, ML-236B (compactin)-producing fungus Penicillium citrinum. Hygromycin B-resistant colonies were obtained after treatment of protoplasts with a vector containing an Escherichia coli hygromycin B phosphotransferase gene fused to a 3-phosphoglycerate kinase promoter from Aspergillus nidulans. The transformation rate was 194 transformants per μg circular DNA per 4x105 viable protoplasts under optimized transformation conditions. Transformation took place via the integration of plasmid DNA into the fungal chromosomal DNA. Most of the integration events appeared to produce tandemly iterated arrays of plasmid molecules at different sites in the chromosome. The transformed, drug-resistant, phenotype and the integrated plasmids were mitotically stable with or without selection in a majority of cases. The demonstration of such a transformation system is an essential first step in the application of recombinant DNA technology to strain improvement and for the production of novel ML-236B derivatives.
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Akins RA, Lambowitz AM (1985) Mol Cell Biol 5:2272–2278
Blochlinger K, Diggelmann H (1984) Mol Cell Biol 4:2929–2931
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Gene 2:95–113
Brown AG, Smale TC, King TJ, Hasenkamp R, Thompson RH (1976) J Chem Soc Perkin Trans 1:1165–1170
Clements JM, Roberts CF (1985) Curr Genet 9:293–298
Clements JM, Roberts CF (1986) Gene 44:97–105
Durand N, Reymond P, Fêvre M (1991) Curr Genet 19:149–153
Endo A, Kuroda M, Tsujita Y (1976a) J Antibiot 29:1346–1348
Endo A, Kuroda M, Tanzawa H (1976b) FEBS Lett 72:323–326
Fincham JRS (1989) Microbiol Rev 53 (1):148–170
Henikoff S (1984) Gene 28:351–359
Hunag D, Bhairi S, Staples RC (1989) Curr Genet 15:411–414
Kaster KR, Burgett SG, Rao RN, Ingolia TD (1983) Nucleic Acids Res 11:6895–6911
Kistler HC, Benny UK (1988) Curr Genet 13:145–149
Nakaya N, Homma Y, Tamachi H, Shigematsu H, Hata Y, Goto Y (1987) J Am Med Assoc 257:3088–3093
Nara F, Watanabe I, Serizawa N (1992) Curr Genet (in press)
Oliver RP, Roberts IN, Harling R, Kenyon L, Punt PJ, Dingemanse MA, van den Hondel CAMJJ (1987) Curr Genet 12:231–233
Punt PJ, Oliver RP, Dingemanse MA, Pouwels PH, van den Hondel CAMJJ (1987) Gene 56:117–124
Queener SW, Ignolia TD, Skatrud PL, Chapman JL, Kaster KR (1985) In: Schlessinger D (ed) Microbiology. Am Soc Microbiol 468–471
Saito Y, Goto Y, Nakaya N, Hata Y, Homma Y, Naito C, Hayashi H, Ito H, Yamamoto M, Takeuchi I, Mori K, Hara T, Yoshida S, Shirai N, Sasaki N, Shinomiya M, Murano S, Morisaki N, Nishiide T, Kanzaki T, Watanabe N, Ishikawa T (1988) Atherosclerosis 72:205–211
Sanger F, Nicklen S, Coulson AR (1977) Proc Natl Acad Sci USA 74:5463–5467
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Serizawa N, Serizawa S, Nakagawa K, Furuya K, Okazaki T, Terahara A (1983) Antibiot 36:887–891
Tilburn J, Scazzocchia C, Taylor GG, Zabicky-Zissman JH, Lockington RA, Davies RW (1983) Gene 26:205–221
Tsujita Y, Kuroda M, Shimada Y, Tanzawa K, Arai M, Kaneko I, Tanaka M, Masuda H, Tarumi C, Watanabe Y, Fujii S (1986) Biochim Biophys Acta 877:50–60
Turner G, Ballance DJ (1985) In: Bennett JW, Lasure LL (eds) Gene manipualtions in fungi. Academic Press, San Diego, pp 259–275
Vogel HJ (1956) Microbiol Genet Bull 13:42–43
Yelton MM, Hamer JE, Timberlake WE (1984) Proc Natl Acid Sci USA 81:1470–1474
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Nara, F., Watanabe, I. & Serizawa, N. Development of a transformation system for the filamentous, ML-236B (compactin) — producing fungus Penicillium citrinum . Curr Genet 23, 28–32 (1993). https://doi.org/10.1007/BF00336746
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DOI: https://doi.org/10.1007/BF00336746