Continuous-exchange cell-free protein synthesis using PCR-generated DNA and an RNase E-deficient extract
DC Field | Value | Language |
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dc.contributor.author | Jun, Soo Youn | - |
dc.contributor.author | Kang, Sang Hyeon | - |
dc.contributor.author | Lee, Kwang-Ho | - |
dc.date.available | 2019-05-30T05:35:38Z | - |
dc.date.issued | 2008-03 | - |
dc.identifier.issn | 0736-6205 | - |
dc.identifier.issn | 1940-9818 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/23835 | - |
dc.description.abstract | Though the use of PCR-generated DNA (i.e., linear template) as template DNA is desirable because of its simple preparation, the linear template has not been routinely used in a conventional continuous-exchange cell-free (CECF) protein synthesis system due to the instability of the linear template and/or its transcript in the relatively long operation period. To overcome this problem and enhance soluble protein yield, an RNase E-deficient and molecular chaperone-enriched extract was used: (i) for compensating for the decrease in messenger RNA (mRNA) levels transcribed from the unstable linear template with improvement of mRNA stability by depletion of RNase E activity; and (ii) for enhancement of the soluble protein portion by assisting of the molecular chaperones. As a result, soluble erythropoietin production from a linear template was significantly enhanced in this modified CECF system using the RNase E-deficient and molecular chaperone-enriched extract, and the amount of soluble erythropoietin was estimated to be roughly 70% of that from a circular plasmid. We can conclude that the use of RNase E-deficient and molecular chaperone-enriched S30 extract mixture is effective in the enhancenment of soluble protein expression from a linear template in the CECF system. | - |
dc.format.extent | 5 | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | FUTURE SCI LTD | - |
dc.title | Continuous-exchange cell-free protein synthesis using PCR-generated DNA and an RNase E-deficient extract | - |
dc.type | Article | - |
dc.identifier.doi | 10.2144/000112690 | - |
dc.identifier.bibliographicCitation | BIOTECHNIQUES, v.44, no.3, pp 387 - 391 | - |
dc.description.isOpenAccess | N | - |
dc.identifier.wosid | 000263637500014 | - |
dc.identifier.scopusid | 2-s2.0-41849151162 | - |
dc.citation.endPage | 391 | - |
dc.citation.number | 3 | - |
dc.citation.startPage | 387 | - |
dc.citation.title | BIOTECHNIQUES | - |
dc.citation.volume | 44 | - |
dc.type.docType | Article | - |
dc.publisher.location | 미국 | - |
dc.subject.keywordPlus | ESCHERICHIA-COLI | - |
dc.subject.keywordPlus | STABILITY | - |
dc.subject.keywordPlus | SYSTEM | - |
dc.subject.keywordPlus | GENES | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Biochemistry & Molecular Biology | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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