Characterisation of the Mucor circinelloides regulated promoter gpd1P.
The promoter of the Mucor circinelloides gpd1 gene encoding glyceraldehyde-3-phosphate dehydrogenase (gpd1P) was recently cloned and used for the production of recombinant proteins, such as the Aspergillus niger glucose oxidase 1 (GOX). This represents the first example of the application of a strong and regulated promoter from this fungus for recombinant protein production. The original 741-bp gpd1P promoter fragment conferred hexose-dependent expression of GOX in M. circinelloides. To understand the regulatory mechanisms involved in gpd1P-driven expression and to develop improved promoter fragments, deletion derivatives of gpd1P were constructed. These derivatives were fused to the A. niger gox1 gene and used to construct strains containing a single copy of the expression cassette. GOX activity was detected in strains containing the full-length gpd1P and also in strains containing a 713-bp or a 361-bp derivative. Expression levels for the 361-bp derivative were high and comparable, regardless of the carbon source used. This promoter represents a useful derivative for constitutive heterologous gene expression in M. circinelloides.
Larsen
Gitte G
GG
Department of Fungal Biotechnology, Biotechnological Institute, Kogle Allé 2, 2970, Hørsholm, Denmark.
Appel
Karen F
KF
Wolff
Anne-Mette
AM
Nielsen
Jens
J
Arnau
José
J
D001483:Base Sequence; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005949:Glucose Oxidase / Q000096:biosynthesis / Q000235:genetics; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D009089:Mucor / Q000235:genetics*; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
D005949:Glucose Oxidase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases