Effects of a minor isoleucyl tRNA on heterologous protein translation in Escherichia coli
10.1128/jb.177.24.7086-7091.1995
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

In Escherichia coli, the isoleucine codon AUA occurs at a frequency of about 0.4% and is the fifth rarest codon in E. coli mRNA. Since there is a correlation between the frequency of codon usage and the level of its cognate tRNA, translational problems might be expected when the mRNA contains high levels of AUA codons. When a hemagglutinin from the influenza virus, a 304-amino-acid protein with 12 (3.9%) AUA codons and 1 tandem codon, and a mupirocin-resistant isoleucyl tRNA synthetase, a 1,024-amino-acid protein, with 33 (3.2%) AUA codons and 2 tandem codons, were expressed in E. coli, product accumulation was highly variable and dependent to some degree on the growth medium. In rich medium, the flu antigen represented about 16% of total cell protein, whereas in minimal medium, it was only 2 to 3% of total cell protein. In the presence of the cloned ileX, which encodes the cognate tRNA for AUA, however, the antigen was 25 to 30% of total cell protein in cells grown in minimal medium. Alternatively, the isoleucyl tRNA synthetase did not accumulate to detectable levels in cells grown in Luria broth unless the ileX tRNA was coexpressed when it accounted for 7 to 9% of total cell protein. These results indicate that the rare isoleucine AUA codon, like the rare arginine codons AGG and AGA, can interfere with the efficient expression of cloned proteins.

Bibliography

Del Tito, B. J., Ward, J. M., Hodgson, J., Gershater, C. J., Edwards, H., Wysocki, L. A., Watson, F. A., Sathe, G., & Kane, J. F. (1995). Effects of a minor isoleucyl tRNA on heterologous protein translation in Escherichia coli. Journal of Bacteriology, 177(24), 7086–7091.

Authors 9
  1. B J Del Tito (first)
  2. J M Ward (additional)
  3. J Hodgson (additional)
  4. C J Gershater (additional)
  5. H Edwards (additional)
  6. L A Wysocki (additional)
  7. F A Watson (additional)
  8. G Sathe (additional)
  9. J F Kane (additional)
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Dates
Type When
Created 8 years, 9 months ago (Nov. 9, 2016, 9:22 a.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:53 p.m.)
Indexed 1 month ago (Aug. 5, 2025, 9:02 a.m.)
Issued 29 years, 9 months ago (Dec. 1, 1995)
Published 29 years, 9 months ago (Dec. 1, 1995)
Published Print 29 years, 9 months ago (Dec. 1, 1995)
Funders 0

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@article{Del_Tito_1995, title={Effects of a minor isoleucyl tRNA on heterologous protein translation in Escherichia coli}, volume={177}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.177.24.7086-7091.1995}, DOI={10.1128/jb.177.24.7086-7091.1995}, number={24}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Del Tito, B J and Ward, J M and Hodgson, J and Gershater, C J and Edwards, H and Wysocki, L A and Watson, F A and Sathe, G and Kane, J F}, year={1995}, month=dec, pages={7086–7091} }