Methods are disclosed for enhancing protein production. One method
comprises preparing a vector by inserting a gene encoding ubiquitin in
front of a gene encoding a protein of interest and inserting the vector
into a cell. A fusion protein will be expressed which includes ubiquitin
plus the protein of interest. Ubiquitin C-terminal hydrolases can cleave
the fusion protein leaving the desired protein in its free state. This
method causes enhanced production of the protein of interest as compared
to performing the same method without the ubiquitin gene as part of the
vector. A ubiquitin promoter is unnecessary to yield this enhanced
production and is not used. A second method is very similar except that
in place of a ubiquitin gene, a gene encoding fourteen amino acids of
cucumber mosaic virus coat protein is inserted in front of the gene of
interest. This results in expression of a fusion protein comprising the
fourteen amino acid residues of the coat protein bonded to the protein of
interest. The fusion protein is produced at a higher level than is the
protein when the coat protein gene fragment is not present in the vector.
In both methods the genes can be placed under the control of heterologous
promoters such as a 35S promoter.