High-level expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene product.
We have observed that proteins, such as human tissue-type plasminogen activator, pro-urokinase or gp41 of human immunodeficiency virus, which have a high content of rare codons in their respective genes, are not readily expressed in Escherichia coli. Furthermore induction of these heterologous genes leads to growth inhibition and plasmid instability. Supplementation with tRNA(AGA/AGG(Arg)) by cotransfection with the dnaY gene, which supplies this minor tRNA, resulted in high-level production with greatly improved cell viability and plasmid stability.
Brinkmann
U
U
Department of Genetics, Boehringer Mannheim GmbH, Penzberg (F.R.G.).
Mattes
R E
RE
Buckel
P
P
D001483:Base Sequence; D004274:DNA, Recombinant / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012347:RNA, Transfer, Arg / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D004274:DNA, Recombinant; D015345:Oligonucleotide Probes; D012345:RNA, Transfer, Amino Acid-Specific; D012347:RNA, Transfer, Arg; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator