title: Translational fusions with fragments of the trpE gene improve the expression of a poorly expressed heterologous gene in Escherichia coli.
abstract: A series of plasmids expressing fusions between the trpE gene product, anthranilate synthase component I and the major immunogen (VP1) of foot and mouth disease virus were constructed such that increasing amounts of the 3' end of trpE were deleted. Deletions removing up to 70% of trpE had little effect on the quantity of fusion protein expressed, while the number of molecules appeared to increase. Larger deletions led to a steady decrease in both the quantity of fusion protein produced and in the number of molecules. This is consistent with trpE being responsible for the high levels of expression of VP1 by its gene product stabilizing VP1 against proteolytic degradation. Some out-of-frame deletion mutants were also produced. All deletion mutants in one wrong reading frame expressed low levels of two VP1-containing polypeptides. This observation is interpreted as being due to re-initiation of translation at a site inside the VP1 sequence which is activated by local termination of translation.
authors.affiliation: Wellcome Biotech Ltd., Kent, UK.
authors.forename: A
authors.initials: A
authors.lastname: Makoff
authors.forename: N
authors.initials: N
authors.lastname: Parry
authors.forename: L
authors.initials: L
authors.lastname: Dicken
mesh_terms: D001483:Base Sequence; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015183:Restriction Mapping
chemical_list: D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins