Expression of a synthetic protein-based polymer (elastomer) gene in Aspergillus nidulans |
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| Authors: | R W Herzog N K Singh D W Urry H Daniell |
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| Institution: | (1) Department of Botany and Microbiology, 101 Life Sciences Building, Auburn University, AL 36849-5407, USA, US;(2) Laboratory of Molecular Biophysics, School of Medicine, The University of Alabama at Birmingham, Birmingham, AL 35294-0019, USA, US |
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| Abstract: | 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.
Received: 23 July 1996 / Received revision: 19 November 1996 / Accepted: 23 November 1996 |
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