Expression of a synthetic protein-based polymer (elastomer) gene in Aspergillus nidulans.
A gene for a synthetic protein-based polymer, G-(VPGVG)119-VPGV, coding for the EG-120mer (elastomer), was cloned into a fungal expression vector to allow constitutive expression of the polymer controlled by the gpdA (glyceraldehyde-3-phosphate dehydrogenase) promoter sequence of Aspergillus nidulans. Stable transformants of A. nidulans showed plasmid integration with varying copy number when analyzed by Southern-blot hybridization. Expression of the synthetic gene was demonstrated by Northern-blot hybridization. However, the translational efficiency for production of the polymer polypeptide was low, presumably because of certain codons in the polymer gene (CCG and GUA) that are rarely used by A. nidulans. Partial purification by reversible phase transition followed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed the presence of polymer protein in a transformant that contained multiple copies of the polymer gene. This study represents the first attempt to express a synthetic gene (with no natural analog) in a fungus.
Herzog
R W
RW
Department of Botany and Microbiology, Auburn University, AL 36849-5407, USA.
Singh
N K
NK
Urry
D W
DW
Daniell
H
H
D001233:Aspergillus nidulans / Q000235:genetics* / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000032:analysis; D012408:Rubber / Q000378:metabolism*; D014170:Transformation, Genetic
D004271:DNA, Fungal; D012331:RNA, Fungal; D012408:Rubber; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases