title: Cell-free translational screening of an expression sequence tag library of Clonorchis sinensis for novel antigen discovery.
abstract: The rapidly evolving cloning and sequencing technologies have enabled understanding of genomic structure of parasite genomes, opening up new ways of combatting parasite-related diseases. To make the most of the exponentially accumulating genomic data, however, it is crucial to analyze the proteins encoded by these genomic sequences. In this study, we adopted an engineered cell-free protein synthesis system for large-scale expression screening of an expression sequence tag (EST) library of Clonorchis sinensis to identify potential antigens that can be used for diagnosis and treatment of clonorchiasis. To allow high-throughput expression and identification of individual genes comprising the library, a cell-free synthesis reaction was designed such that both the template DNA and the expressed proteins were co-immobilized on the same microbeads, leading to microbead-based linkage of the genotype and phenotype. This reaction configuration allowed streamlined expression, recovery, and analysis of proteins. This approach enabled us to identify 21 antigenic proteins. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:832-837, 2017.
authors.affiliation: Dept. of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea.
authors.forename: Devi
authors.initials: D
authors.lastname: Kasi
authors.affiliation: Dept. of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea.
authors.forename: Christy
authors.initials: C
authors.lastname: Catherine
authors.affiliation: Dept. of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea.
authors.forename: Seung-Won
authors.initials: SW
authors.lastname: Lee
authors.affiliation: Dept. of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea.
authors.forename: Kyung-Ho
authors.initials: KH
authors.lastname: Lee
authors.affiliation: Div. of Malaria and Parasitic Diseases, National Institute of Health, Osong, 361-951, Korea.
authors.forename: Yu Jung
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Div. of Malaria and Parasitic Diseases, National Institute of Health, Osong, 361-951, Korea.
authors.forename: Myeong
authors.initials: M
authors.lastname: Ro Lee
authors.affiliation: Div. of Malaria and Parasitic Diseases, National Institute of Health, Osong, 361-951, Korea.
authors.forename: Jung Won
authors.initials: JW
authors.lastname: Ju
authors.affiliation: Dept. of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon, 305-764, Korea.
authors.forename: Dong-Myung
authors.initials: DM
authors.lastname: Kim
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology* / Q000378:metabolism; D003003:Clonorchiasis / Q000276:immunology* / Q000469:parasitology; D003004:Clonorchis sinensis / Q000276:immunology* / Q000378:metabolism / Q000472:pathogenicity; D019295:Computational Biology; D020224:Expressed Sequence Tags; D005838:Genotype; D010641:Phenotype; D040901:Proteomics
chemical_list: D000941:Antigens
keywords: Clonorchis sinensis; bead display; cell-free protein synthesis; high-throughput expression; parasite antigen