In silico analysis of chimeric espA, eae and tir fragments of Escherichia coli O157:H7 for oral immunogenic applications.
BACKGROUND
In silico techniques are highly suited for both the discovery of new and development of existing vaccines. Enterohemorrhagic Escherichia coli O157:H7 (EHEC) exhibits a pattern of localized adherence to host cells, with the formation of microcolonies, and induces a specific histopathological lesion (attaching/effacing). The genes encoding the products responsible for this phenotype are clustered on a 35-kb pathogenicity island. Among these proteins, Intimin, Tir, and EspA, which are expressed by attaching-effacing genes, are responsible for the attachment to epithelial cell that leads to lesions.


RESULTS
We designed synthetic genes encoding the carboxy-terminal fragment of Intimin, the middle region of Tir and the carboxy-terminal part of EspA. These multi genes were synthesized with codon optimization for a plant host and were fused together by the application of four repeats of five hydrophobic amino acids as linkers. The structure of the synthetic construct gene, its mRNA and deduced protein and their stabilities were analyzed by bioinformatic software. Furthermore, the immunogenicity of this multimeric recombinant protein consisting of three different domains was predicted.


CONCLUSIONS
a structural model for a chimeric gene from LEE antigenic determinants of EHEC is presented. It may define accessibility, solubility and immunogenecity.
Amani
Jafar
J
National Institute of Genetic Engineering and Biotechnology, Shahrak-e- Pajoohesh, 15th Km, Tehran -Karaj Highway, Tehran, IR, Iran. jamani@nigeb.ac.ir
Mousavi
S Latif
SL
Rafati
Sima
S
Salmanian
Ali H
AH
D018829:Adhesins, Bacterial / Q000737:chemistry; D001422:Bacterial Adhesion; D003062:Codon; D019295:Computational Biology / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D019453:Escherichia coli O157 / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry; D044404:Genomic Islands; D008962:Models, Theoretical; D010641:Phenotype; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D018829:Adhesins, Bacterial; D003062:Codon; D029968:Escherichia coli Proteins; C102051:EspA protein, E coli; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C109287:Tir protein, E coli; C073819:eaeA protein, E coli