Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates.
Progress towards genetic sequencing of human parasites has provided the groundwork for a post-genomic approach to develop novel antigens for the diagnosis and treatment of parasite infections. To fully utilize the genomic data, however, high-throughput methodologies are required for functional analy...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4657965?pdf=render |
| _version_ | 1818269984938262528 |
|---|---|
| author | Christy Catherine Seung-Won Lee Jung Won Ju Ho-Cheol Kim Hyun-Il Shin Yu Jung Kim Dong-Myung Kim |
| author_facet | Christy Catherine Seung-Won Lee Jung Won Ju Ho-Cheol Kim Hyun-Il Shin Yu Jung Kim Dong-Myung Kim |
| author_sort | Christy Catherine |
| collection | DOAJ |
| description | Progress towards genetic sequencing of human parasites has provided the groundwork for a post-genomic approach to develop novel antigens for the diagnosis and treatment of parasite infections. To fully utilize the genomic data, however, high-throughput methodologies are required for functional analysis of the proteins encoded in the genomic sequences. In this study, we investigated cell-free expression and in situ immobilization of parasite proteins as a novel platform for the discovery of antigenic proteins. PCR-amplified parasite DNA was immobilized on microbeads that were also functionalized to capture synthesized proteins. When the microbeads were incubated in a reaction mixture for cell-free synthesis, proteins expressed from the microbead-immobilized DNA were instantly immobilized on the same microbeads, providing a physical linkage between the genetic information and encoded proteins. This approach of in situ expression and isolation enables streamlined recovery and analysis of cell-free synthesized proteins and also allows facile identification of the genes coding antigenic proteins through direct PCR of the microbead-bound DNA. |
| first_indexed | 2024-12-12T21:03:04Z |
| format | Article |
| id | doaj.art-f0005b63955c481fb1b8465d69c8010d |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T21:03:04Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f0005b63955c481fb1b8465d69c8010d2022-12-22T00:12:06ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011011e014359710.1371/journal.pone.0143597Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates.Christy CatherineSeung-Won LeeJung Won JuHo-Cheol KimHyun-Il ShinYu Jung KimDong-Myung KimProgress towards genetic sequencing of human parasites has provided the groundwork for a post-genomic approach to develop novel antigens for the diagnosis and treatment of parasite infections. To fully utilize the genomic data, however, high-throughput methodologies are required for functional analysis of the proteins encoded in the genomic sequences. In this study, we investigated cell-free expression and in situ immobilization of parasite proteins as a novel platform for the discovery of antigenic proteins. PCR-amplified parasite DNA was immobilized on microbeads that were also functionalized to capture synthesized proteins. When the microbeads were incubated in a reaction mixture for cell-free synthesis, proteins expressed from the microbead-immobilized DNA were instantly immobilized on the same microbeads, providing a physical linkage between the genetic information and encoded proteins. This approach of in situ expression and isolation enables streamlined recovery and analysis of cell-free synthesized proteins and also allows facile identification of the genes coding antigenic proteins through direct PCR of the microbead-bound DNA.http://europepmc.org/articles/PMC4657965?pdf=render |
| spellingShingle | Christy Catherine Seung-Won Lee Jung Won Ju Ho-Cheol Kim Hyun-Il Shin Yu Jung Kim Dong-Myung Kim Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. PLoS ONE |
| title | Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. |
| title_full | Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. |
| title_fullStr | Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. |
| title_full_unstemmed | Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. |
| title_short | Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates. |
| title_sort | cell free expression and in situ immobilization of parasite proteins from clonorchis sinensis for rapid identification of antigenic candidates |
| url | http://europepmc.org/articles/PMC4657965?pdf=render |
| work_keys_str_mv | AT christycatherine cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT seungwonlee cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT jungwonju cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT hocheolkim cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT hyunilshin cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT yujungkim cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates AT dongmyungkim cellfreeexpressionandinsituimmobilizationofparasiteproteinsfromclonorchissinensisforrapididentificationofantigeniccandidates |