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Purification of an Intact Human Protein Overexpressed from Its Endogenous Locus via Direct Genome Engineering
DC Field | Value | Language |
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dc.contributor.author | 조성래 | - |
dc.date.accessioned | 2020-09-29T01:11:21Z | - |
dc.date.available | 2020-09-29T01:11:21Z | - |
dc.date.issued | 2020-06 | - |
dc.identifier.uri | https://ir.ymlib.yonsei.ac.kr/handle/22282913/179429 | - |
dc.description.abstract | The overproduction and purification of human proteins is a requisite of both basic and medical research. Although many recombinant human proteins have been purified, current protein production methods have several limitations; recombinant proteins are frequently truncated, fail to fold properly, and/or lack appropriate post-translational modifications. In addition, such methods require subcloning of the target gene into relevant plasmids, which can be difficult for long proteins with repeated domains. Here we devised a novel method for target protein production by introduction of a strong promoter for overexpression and an epitope tag for purification in front of the endogenous human gene, in a sense performing molecular cloning directly in the human genome, which does not require cloning of the target gene. As a proof of concept, we successfully purified intact human Reelin protein, which is lengthy (3460 amino acids) and contains repeating domains, and confirmed that it was biologically functional. | - |
dc.description.statementOfResponsibility | restriction | - |
dc.language | English | - |
dc.publisher | American Chemical Society | - |
dc.relation.isPartOf | ACS SYNTHETIC BIOLOGY | - |
dc.rights | CC BY-NC-ND 2.0 KR | - |
dc.title | Purification of an Intact Human Protein Overexpressed from Its Endogenous Locus via Direct Genome Engineering | - |
dc.type | Article | - |
dc.contributor.college | College of Medicine (의과대학) | - |
dc.contributor.department | Dept. of Rehabilitation Medicine (재활의학교실) | - |
dc.contributor.googleauthor | Jihyeon Yu | - |
dc.contributor.googleauthor | Eunju Cho | - |
dc.contributor.googleauthor | Yeon-Gil Choi | - |
dc.contributor.googleauthor | You Kyeong Jeong | - |
dc.contributor.googleauthor | Yongwoo Na | - |
dc.contributor.googleauthor | Jong-Seo Kim | - |
dc.contributor.googleauthor | Sung-Rae Cho | - |
dc.contributor.googleauthor | Jae-Sung Woo | - |
dc.contributor.googleauthor | Sangsu Bae | - |
dc.identifier.doi | 10.1021/acssynbio.0c00090 | - |
dc.contributor.localId | A03831 | - |
dc.relation.journalcode | J03860 | - |
dc.identifier.eissn | 2161-5063 | - |
dc.identifier.pmid | 32584551 | - |
dc.identifier.url | https://pubs.acs.org/doi/10.1021/acssynbio.0c00090 | - |
dc.subject.keyword | CRISPR-Cas system | - |
dc.subject.keyword | Reelin | - |
dc.subject.keyword | genome engineering | - |
dc.subject.keyword | molecular cloning | - |
dc.subject.keyword | protein purification | - |
dc.contributor.alternativeName | Cho, Sung Rae | - |
dc.contributor.affiliatedAuthor | 조성래 | - |
dc.citation.volume | 9 | - |
dc.citation.number | 7 | - |
dc.citation.startPage | 1591 | - |
dc.citation.endPage | 1598 | - |
dc.identifier.bibliographicCitation | ACS SYNTHETIC BIOLOGY, Vol.9(7) : 1591-1598, 2020-06 | - |
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