Isolation and characterization of the 3-phosphoglycerate kinase gene (pgk) from the filamentous fungus Trichoderma reesei

Abstract

The 3-phosphoglycerate kinase gene (pgk) from Trichoderma reesei was isolated by hybridization with the corresponding Saccharomyces cerevisiae PGK gene. The 1,545 nt long nucleotide sequence of the cloned gene codes for a 416 amino acid protein. The coding sequence contains two introns of 219 and 75 nt, respectively, at positions identical to those corresponding genes from the other filamentous fungi Aspergillus nidulans and Penicillum chrysogenum. This gene codes for two mRNAs of about 1.65 kb and 1.85 kb. The PGK protein of Trichoderma shows extensive homology to the PGKs of other fungi A. nidulans (77%), P. chrysogenum (73%) and Saccharomyces cerevisiae (69%). However, the PGKs of the two other filamentous fungi, A. nidulans and P. chrysogenum, seem to be more closely related to each other than to the T. reesei enzyme.

Differential Expression of the Vegetative and Spore‐Bound Hydrophobins of Trichoderma Reesei Cloning and Characterization of the Hfb2 Gene

“…The chromosomal copy of the gene was isolated from a 7: reesei QM9414 genomic i. bank (Vanhanen et al, 1989) by using the cDNA clone as a probe. The chromosomal gene was subcloned on the basis of Southern analysis of restriction enzyme digestions carried out for the isolated 2 clones.…”

“…12 clones giving a weak hybridization signal with the low stringent washes but no signal with the high stringent washes were picked, rescreened, and in vivo excised into phagemid pBluescriptSK( -) form as described by the manufacturer, resulting in clones phfb2/l -phfb2/12. The hfb2 genomic copy was isolated from a 7: reesei QM9414 genomic lambda-library (Vanhanen et al, 1989) by using as a probe a [a-…”

Overexpression of bacterial ethylene-forming enzyme gene in Trichoderma reesei enhanced the production of ethylene

“…In the expression vector pPpgk1-efe-hph, the efe gene was controlled by a strong constitutive pgk I promoter from T. reesei 19,20. This promoter was amplified from the genomic DNA of T. reesei QM9414 using PCR with P7 primer (5'-ACAGCATGCGATGATGGAGGATATACGCGA-3'), including an engineered Sph I site; P8 primer (5'-ACGGTCGACGTTAGACAGAGACATTTTGGC-3'), including an engineered Sal I site.…”

Metabolic Diversity of Trichoderma

“…The genes encoding 3-phosphoglycerate kinase and pyruvate kinase from T. reesei and T. viride have been cloned (Goldman et al, 1992;Schindler et al, 1993;Vanhanen et al, 1989). The promoter region of 3-phosphoglycerate kinase has consensus binding sites for a cyclic-AMP responsive element and the carbon catabolite repressor Cre1 (Vanhanen et al, 1991).…”