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KMID : 1034820170130040387
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Molecular & Cellular Toxicology
2017 Volume.13 No. 4 p.387 ~ p.394
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Heterogeneous expression of Chlamydia pneumoniae antigen candidates and high-level soluble expression of its inclusion membrane proteins in Escherichia coli
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Lee Soo-Youn
Lee Sun-Jin
Ahn Ji-Young
Ko Jung-Ho
Lee Lyon
Han Janet
Kim Young-Chang
Kim Yang-Hoon
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Abstract
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Chlamydia pneumoniae is an intracellular bacterium associated with several acute respiratory diseases and is an obligate intracellular parasite in eukaryotic cells. Heterogeneous and overexpression of virulence-associated C. pneumoniae “effector” proteins with structurally designed epitope-focused antigens in recombinant forms in Escherichia coli is a prerequisite in the study of the organism’s pathogenic mechanisms and for use in medical applications such as vaccine and therapeutics development. In this study, seven antigen candidates from C. pneumoniae TW-183 (OmpH, Omp11, Eno, ArtJ, IncA, IncB and IncC) were screened in silico genome. Immunogenicity of the heterogeneously overexpressed antigen candidates in E. coli BL21 (DE3) was investigated by Western blot analysis using in sera from C. pneumoniae infected individuals. The soluble forms of three inclusion membrane proteins (IncA, IncB and IncC), carrying unique bilobal hydrophobic domains, were also successfully expressed in E. coli BL21 (DE3) through truncation of their membrane bound transmembrane domains.
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KEYWORD
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Chlamydia pneumoniae, Antigen candidates, Heterogeneous expression, Inclusion membrane protein, Immunogenicity
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