Vv. Joutsjoki, CONSTRUCTION BY ONE-STEP GENE REPLACEMENT OF TRICHODERMA-REESEI STRAINS THAT PRODUCE THE GLUCOAMYLASE-P OF HORMOCONIS-RESINAE, Current genetics, 26(5-6), 1994, pp. 422-429
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 cell
obiohydrolase 1 gene (cbh1), were constructed and used 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 sequen
ces in the genomic gamP gene are processed in T. reesei. According to
the results, a T. reesei transformant strain, in which the cbh1 gene h
as been replaced by a single copy of the gamP genomic gene, secretes m
ore active GAMP than does a transformant strain having three copies of
the cDNA clone in tandem orientation at the cbh1 locus.