dc.contributor.author | Ahn, Jin Ho | |
dc.contributor.author | Keum, Jung-Won | |
dc.contributor.author | Kim, Dong-Myung | |
dc.date.accessioned | 2012-02-08T20:12:01Z | |
dc.date.available | 2012-02-08T20:12:01Z | |
dc.date.issued | 2011-11 | |
dc.date.submitted | 2011-04 | |
dc.identifier.issn | 1932-6203 | |
dc.identifier.uri | http://hdl.handle.net/1721.1/69049 | |
dc.description.abstract | While access to soluble recombinant proteins is essential for a number of proteome studies, preparation of purified functional proteins is often limited by the protein solubility. In this study, potent solubility-enhancing fusion partners were screened from the repertoire of endogenous E. coli proteins. Based on the presumed correlation between the intracellular abundance and folding efficiency of proteins, PCR-amplified ORFs of a series of highly abundant E. coli proteins were fused with aggregation-prone heterologous proteins and then directly expressed for quantitative estimation of the expression efficiency of soluble translation products. Through two-step screening procedures involving the expression of 552 fusion constructs targeted against a series of cytokine proteins, we were able to discover a number of endogenous E. coli proteins that dramatically enhanced the soluble expression of the target proteins. This strategy of cell-free expression screening can be extended to quantitative, global analysis of genomic resources for various purposes. | en_US |
dc.description.sponsorship | National Research Foundation of Korea | en_US |
dc.description.sponsorship | Korea (South). Ministry of Education, Science and Technology (MEST) (grant 2011K000841) | en_US |
dc.description.sponsorship | Korea (South). Ministry of Education, Science and Technology (MEST) (grant 2011-0027901) | en_US |
dc.language.iso | en_US | |
dc.publisher | Public Library of Science | en_US |
dc.relation.isversionof | http://dx.doi.org/10.1371/journal.pone.0026875 | en_US |
dc.rights | Creative Commons Attribution | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by/2.5/ | en_US |
dc.source | PLoS | en_US |
dc.title | Expression Screening of Fusion Partners from an E. coli Genome for Soluble Expression of Recombinant Proteins in a Cell-Free Protein Synthesis System | en_US |
dc.type | Article | en_US |
dc.identifier.citation | Ahn, Jin-Ho, Jung-Won Keum, and Dong-Myung Kim. “Expression Screening of Fusion Partners from an E. coli Genome for Soluble Expression of Recombinant Proteins in a Cell-Free Protein Synthesis System.” Ed. Anna Mitraki. PLoS ONE 6.11 (2011): e26875. Web. 8 Feb. 2012. | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Department of Chemical Engineering | en_US |
dc.contributor.approver | Ahn, Jin Ho | |
dc.contributor.mitauthor | Ahn, Jin Ho | |
dc.relation.journal | PLoS ONE | en_US |
dc.eprint.version | Final published version | en_US |
dc.type.uri | http://purl.org/eprint/type/JournalArticle | en_US |
eprint.status | http://purl.org/eprint/status/PeerReviewed | en_US |
dspace.orderedauthors | Ahn, Jin-Ho; Keum, Jung-Won; Kim, Dong-Myung | en |
mit.license | PUBLISHER_CC | en_US |
mit.metadata.status | Complete | |