ArticleMicrobial cell factories2017
Effects of mRNA secondary structure on the expression of HEV ORF2 proteins in Escherichia coli.
Article in Microbial cell factories, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 30 citations in OpenAlex.
- Design of a novel multi epitope antigen for diagnosis of visceral leishmaniasis using an immunoinformatics approach.Scientific reports · 2025Article
- Thermostable Esterase from ThermophilicGenes · 2025Article
- Recent Advances in mRNA-Based Vaccines Against Several Hepatitis Viruses.Biological procedures online · 2025Review
- Decoding stoichiometric protein synthesis inBiophysical reports · 2023Article
- Design and development of a chimeric vaccine candidate against zoonotic hepatitis E and foot-and-mouth disease.Microbial cell factories · 2020Article
- High-resolution modeling of the selection on local mRNA folding strength in coding sequences across the tree of life.Genome biology · 2020Article
- Development of BacMam Induced Hepatitis E Virus Replication Model in Hepatoma Cells to Study the Polyprotein Processing.Frontiers in microbiology · 2020Article
- Development of new hepatitis E vaccines.Human vaccines & immunotherapeutics · 2018Review
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundViral protein expression in Escherichia coli (E. coli) is a powerful tool for structural/functional studies as well as for vaccine and diagnostics development. However, numerous factors such as codon bias, mRNA secondary structure and nucleotides distribution, have been indentified to hamper this heterologous expression.
resultsIn this study, we combined computational and biochemical methods to analyze the influence of these factors on the expression of different segments of hepatitis E virus (HEV) ORF 2 protein and hepatitis B virus surface antigen (HBsAg). Three out of five HEV antigens were expressed while all three HBsAg fragments were not. The computational analysis revealed a significant difference in nucleotide distribution between expressed and non-expressed genes; and all these non-expressing constructs shared similar stable 5'-end mRNA secondary structures that affected the accessibility of both Shine-Dalgarno (SD) sequence and start codon AUG. By modifying the 5'-end of HEV and HBV non-expressed genes, there was a significant increase in the total free energy of the mRNA secondary structures that permitted the exposure of the SD sequence and the start codon, which in turn, led to the successful expression of these genes in E. coli.
conclusionsThis study demonstrates that the mRNA secondary structure near the start codon is the key limiting factor for an efficient expression of HEV ORF2 proteins in E. coli. It describes also a simple and effective strategy for the production of viral proteins of different lengths for immunogenicity/antigenicity comparative studies during vaccine and diagnostics development.
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