title: Expression of a Chimeric Allergen with High Rare Codons Content in Codon Bias-Adjusted Escherichia coli: Escherichia coli BL21 (DE3)-Codon Plus RIL as an Efficient Host.
abstract: The expression of heterologous proteins in Escherichia coli (E. coli) is importantly affected by codon bias. Hence, the aim of the current study was to determine which codon bias-adjusted E. coli strain is sufficient for expression of a chimeric allergen coded by high rare codon content. To investigate the expression level, a chimeric protein of Chenopodium album (C. album) was used as an appropriate model. An expression construct was assembled and was transformed to four strains of codon bias-adjusted E. coli including origami, BL21 (DE3), BL21 (DE3)-codon plus RIL, and Rosetta. The level of expression and solubility of the chimeric allergen was analyzed by SDS-PAGE. In addition, the allergenicity of chimeric allergen was determined using immunoblotting. Our results showed that the chimeric allergen was expressed at high level in E. coli BL21 (DE3)-codon plus RIL and Rosetta. In detail, this recombinant allergen was isolated from soluble fraction in the codon bias-adjusted strains of E. coli BL21 (DE3)-codon plus RIL and Rosetta. Moreover, some lower molecular weight proteins were observed in Rosetta, which could be related to inappropriate expression or broken compartments of the chimeric allergen. The immunoblotting assay confirmed that the IgE-specific immune reactivity of our chimeric allergen expressed in BL21 (DE3)-codon plus RIL was significantly higher than the other strains. Our results showed that the expression of the chimeric allergen with high rare codons content in a codon bias-adjusted strain E. coli BL21 (DE3)-codon plus RIL improves the quality and solubility of the heterologous protein production.
authors.affiliation: Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.
authors.forename: Hamid Reza
authors.initials: HR
authors.lastname: Nouri
authors.affiliation: Student Research Committee, School of Medicine, Babol University of Medical Sciences, Babol, Iran.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Karkhah
authors.affiliation: Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Abdolreza
authors.initials: A
authors.lastname: Varasteh
authors.affiliation: Immunobiochemistry Lab, Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. sankianm@mums.ac.ir.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000378:metabolism; D052179:Antigens, Plant / Q000235:genetics* / Q000378:metabolism; D027463:Chenopodium album / Q000235:genetics*; D003062:Codon / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D010940:Plant Proteins / Q000235:genetics*; D015202:Protein Engineering
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D003062:Codon; D010940:Plant Proteins