Abstract
Two one-step gene replacement vectors containing either the Hormoconis resinae glucoamylase P (gamP) genomic gene or the corresponding cDNA, each under the control of the promoter of the Trichoderma reesei cellobiohydrolase 1 gene (cbh1), were constructed and use to replace the cbh1 gene in a T. reesei strain. In both vectors the cbh1 promoter is precisely fused to the gamP protein coding region. Both the gamP cDNA and the genomic gene direct the secretion of the active glucoamylase P (GAMP) enzyme from T. reesei, which indicates that the intron sequences in the genomic gamP gene are processed in T. reesei. According to the results, a T. reesei transformant strain, in which the cbh1 gene has been replaced by a single copy of the gamP genomic gene, secretes more active GAMP than does a transformant strain having three copies of the cDNA clone in tandem orientation at the cbh1 locus.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aho S, Olkkonen V, Jalava T, Paloheimo M, Bühler R, Niku-Paavola M-L, Bamford DH, Korhola M (1991) Monoclonal antibodies against core and cellulase-binding domains of Trichoderma reesei cellobiohydrolases I and II and endoglucanase I. Eur J Biochem 200:643–649
Bailey MJ, Nevalainen KMH (1981) Induction, isolation and testing of stable Trichoderma reesei mutants with improved production of solubilizing cellulase. Enzyme Microbiol Technol 3:153–157
Banerji J, Olson J, Schaffner W (1983) A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell 33:729–740
Berka RM, Ward M, Wilson LJ, Hayenga KJ, Kodama KH, Carlomagno LP, Thompson SA (1990) Molecular cloning and deletion of the gene encoding aspergillopepsin A from Aspergillus awamori. Gene 86:153–162
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acad Res 7:1513–1523
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Durand H, Clanet M, Tiraby G (1988) Genetic improvement of Trichoderma reesei for large scale cellulase production. Enzyme Microbiol Technol 10:341–346
Esser K, Mohr G (1986) Integrative transformation of filamentous fungi with resepct to biotechnological application. Process Biochem Oct:153–159
Fagerström R (1994) Purification and specificity of recombinant Hormoconis resinae glucoamylase P and endogenous glucoamylase from Trichoderma reesei. Enzyme Microbiol Technol 16:36–42
Fagerström R, Vainio A, Suoranta K, Pakula T, Kalkkinen N, Torkkeli H (1990) Comparison of two glucoamylases from Hormoconis resinae. J Gen Microbiol 136:913–920
Finkelstein DB (1992) Transformation. In: Finkelstein DB, Ball C (eds) Biotechnology of filamentous fungi. Technology and products. Butterworth-Heinemann, Boston, pp 113–156
Harkki A, Uusitalo J, Bailey M, Penttilä M, Knowles JKC (1989) A novel fungal expression system: secretion of active calf chymosin from the filamentous fungus Trichoderma reesei. Bio/Technology 7:596–603
Hinnen A, Hicks JB, Fink GR (1978) Transformation of yeast. Proc Natl Acad Sci USA 75:1929–1933
Joutsjoki VV, Torkkeli TK (1992) Glucoamylase P gene of Hormoconis resinae: molecular cloning, sequencing and introduction into Trichoderma reesei. FEMS Microbiol Lett 99:237–244
Joutsjoki VV, Torkkeli TK, Nevalainen KMH (1993) Transformation of Trichoderma reesei with the Hormoconis resinae glucoamylase P (gamP) gene: production of a heterologous glucoamylase by Trichoderma reesei. Curr Genet 24:223–228
Karhunen T, Mäntylä A, Nevalainen KMH, Suominen PL (1993) High frequency one-step gene replacement in Trichoderma reesei. I. Endoglucanase I overproduction. Mol Gen Genet 241:515–522
Kelly JM, Hynes MJ (1985) Transformation of Aspergillus niger by the amdS gene of Aspergillus nidulans. EMBO J 4:475–479
Konieczny SF, Emerson Jr CP (1987) Complex regulation of the muscle-specific contractile protein (troponin I) gene. Mol Cell Biol 7:3065–3075
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Miller BL, Miller KY, Timberlake WE (1985) Direct and indirect gene replacements in Aspergillus nidulans. Mol Cell Biol 5:1714–1721
Nevalainen H, Penttilä M, Harkki A, Teeri T, Knowles JKC (1991) The molecular biology of Trichoderma and its application to the expression of both homologous and heterologous gene. In: Leong SA, Berka R (eds) Molecular industrial mycology. Systems and applications for filamentous fungi. Marcel Dekker Inc, New York, pp 129–148
Orr-Weaver TL, Szostack JW, Rothstein RJ (1981) Yeast transformantion: a model system for the study of recombination. Proc Natl Acad Sci USA 78:6354–6358
Penttilä M, Nevalainen H, Rättö M, Salminen E, Knowles J (1987) A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei. Gene 61:155–164
Prentice N (1982) Purification of beta-glucanase for beta-D-glucan assays. Cereal Chem 59:231–232
Raeder U, Broda P (1985) Rapid preparation of DNA from filamentous fungi. Lett Appl Microbiol 1:17–20
Rossi P, de Crombrugghe B (1987) Identification of a cell-specific transcriptional enhancer in the first intron of the mouse α2 (type I) collagen gene. Proc Natl Acad Sci USA 84:5590–5594
Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211
Saloheimo M, Niku-Paavola M-L (1991) Heterologous production of a ligninolytic enzyme: expression of the Phlebia radiata laccase gene in Trichoderma reesei. Bio/Technology 9:987–990
Scherer S, Davis RW (1979) Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci USA 76:4951–4955
Seiboth B, Messner R, Gruber F, Kubicek CP (1992) Disruption of the Trichoderma reesei cbh2 gene coding for cellobiohydrolase II leads to a delay in the triggering of cellulase formation by cellulose. J Gen Microbiol 138: 1259–1264
Shoemaker S, Schweickart V, Ladner M, Gelfand D, Kwok S, Myambo K, Innis M (1983) Molecular cloning of exo-cellobiohydrolase I derived from Trichoderma reesei strain L27. Bio/Technology 1:691–696
Slater EP, Rabenau O, Karin M, Baxter JD, Beato M (1985) Glucocorticoid receptor binding and activation of a heterologous promoter by dexamethasone by the first intron of the human growth hormone gene. Mol Cell Biol 5:2984–2992
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Suominen P, Mäntylä A, Saarelainen R, Paloheimo M, Fagerström R, Parkkinen E, Nevalainen H (1992) Genetic engineering of Trichoderma reesei to produce suitable enzyme combinations for applications in the pulp and paper industry. In: Kuwahara M, Shimada M (eds) Biotechnology in pulp and paper industry. Uni Publishers Co, Ltd, Tokyo, Japan, pp 439–445
Suominen PL, Mäntylä AL, Karhunen T, Hakola S, Nevalainen H (1993) High-frequency one-step gene replacement in Trichoderma reesei. II. Effects of deletions of individual cellulase genes Mol Gen Genet 241: 523–530
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Vainio AEI, Torkkeli HT, Tuusa T, Aho SA, Fagerström BR, Korhola MP (1993) Cloning and expression of Hormoconis resinae glucoamylase P cDNA in Saccharomyces cerevisiae. Curr Genet 24: 38–44
Author information
Authors and Affiliations
Additional information
Communicated by A. Hinnen
Rights and permissions
About this article
Cite this article
Joutsjoki, V.V. Construction by one-step gene replacement of Trichoderma reesei strains that produce the glucoamylase P of Hormoconis resinae . Curr Genet 26, 422–429 (1994). https://doi.org/10.1007/BF00309929
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00309929