Efficient GFP expression in the mushrooms Agaricus bisporus and Coprinus cinereus requires introns.
We have developed a "Molecular Toolkit" comprising interchangeable promoters and marker genes to facilitate transformation of homobasidiomycete mushrooms. We describe the evaluation of a range of promoters in the homobasidiomycetes Agaricus bisporus and Coprinus cinereus using green fluorescent protein (GFP) as a reporter gene; the C. cinereus trp1 promoter and A. bisporus trp2 and gpdII promoters proving successful in driving expression in C. cinereus, with the gpdII promoter also functioning in A. bisporus. Our investigations demonstrate that a prerequisite for GFP expression in C. cinereus and A. bisporus is the presence of an intron. This is the first reported expression of GFP in either C. cinereus or A. bisporus.
Burns
C
C
School of Biological Sciences, University of Bristol, Woodland Road, Bristol BS8 1UG, UK.
Gregory
K E
KE
Kirby
M
M
Cheung
M K
MK
Riquelme
M
M
Elliott
T J
TJ
Challen
M P
MP
Bailey
A
A
Foster
G D
GD
D000364:Agaricus / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003302:Coprinus / Q000201:enzymology* / Q000235:genetics; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D007438:Introns / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D015183:Restriction Mapping
D017931:DNA Primers; D004271:DNA, Fungal; D049452:Green Fluorescent Proteins