Cloning and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene and the use of its promoter for expression in Myrothecium gramineum, a novel expression host.
At our laboratory, research has focused on the development of Myrothecium gramineum as a novel expression host. The glyceraldehyde-3-phosphate dehydrogenase (gpd)-promoter of M. gramineum was isolated and characterized (Genbank accession number EF486690). In order to prove its functionality and to explore the potential of M. gramineum as a novel fungal expression host, use of this gpd-promoter for the expression of a fungal alpha-amylase was investigated. Myrothecium gramineum was transformed with pGPDlpAmyAO, containing the gpd-promoter followed by the amy3 encoding sequence of Aspergillus oryzae. Study of the amylase production indicated that the promoter can be successfully used for the expression of heterologous proteins in M. gramineum. To the best of our knowledge, this is the first time a homologous expression system has been described for M. gramineum.
De Maeseneire
Sofie L
SL
Department of Biochemical and Microbial Technology, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium. sofie.demaeseneire@UGent.be
Dauvrin
Thierry
T
Jonniaux
Jean-Luc
JL
Van Bogaert
Inge N A
IN
Soetaert
Wim K
WK
Vandamme
Erick J M C
EJ
D000595:Amino Acid Sequence; D000681:Amylases / Q000235:genetics / Q000378:metabolism; D001236:Aspergillus oryzae / Q000201:enzymology; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D018628:Gene Dosage; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics*; D006999:Hypocreales / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000681:Amylases