Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon
DC Field | Value | Language |
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dc.contributor.author | 이성혁 | - |
dc.contributor.author | 김민식 | - |
dc.contributor.author | 정해창 | - |
dc.contributor.author | 이진원 | - |
dc.contributor.author | 이정현 | - |
dc.contributor.author | 이현숙 | - |
dc.contributor.author | 강성균 | - |
dc.date.accessioned | 2020-07-16T01:52:41Z | - |
dc.date.available | 2020-07-16T01:52:41Z | - |
dc.date.created | 2020-02-11 | - |
dc.date.issued | 2015-04-16 | - |
dc.identifier.uri | https://sciwatch.kiost.ac.kr/handle/2020.kiost/25644 | - |
dc.description.abstract | A strong promoter increases transcription of the genes of interest and enhances the production of various valuable substances. For a hyperthermophilic archaeon Thermococcus onnurineus NA1, which can produce H2 by carbon monoxide oxidation, we searched for a novel endogenous strong promoter by transcriptome analysis using high-throughput RNA sequencing. Based on the relative transcript abundance, we selected one promoter to encode a hypothetical gene, of which homologs were found only in several Thermococcales strains. This promoter, PTN0510, was introduced into the front of CO-responsible hydrogenase gene cluster encoding a carbon monoxide dehydrogenase (CODH), a hydrogenase and a Na+/H+ antiporter. In the resulting mutant strain, KS0510, transcription and translation level of the gene cluster increased by 4- to 14-folds and 1.5- to 1.9-folds, respectively, in comparison with those of wild-type strain. Additionally, H2 production rate of KS0510 mutant was 4.8-fold higher than that of wild-type strain. The PTN0510 was identified to be much stronger than the well-known two strong promoters, gdh and slp promoters from Thermococcus strains, through RT-qPCR and western blotting analyses and kinetics of H2 production. In this study, we demonstrated that the RNA-seq approach is a good strategy to mine a novel strong promoter of use to a Thermococcus strain when developed as a biotechnologically promising strain to produce vwe searched for a novel endogenous strong promoter by transcriptome analysis using high-throughput RNA sequencing. Based on the relative transcript abundance, we selected one promoter to encode a hypothetical gene, of which homologs were found only in several Thermococcales strains. This promoter, PTN0510, was introduced into the front of CO-responsible hydrogenase gene cluster encoding a carbon monoxide dehydrogenase (CODH), a hydrogenase and a Na+/H+ antiporter. In the resulting mutant strain, KS0510, transcription and translation level of the gene cluster increased by 4- to 14-folds and 1.5- to 1.9-folds, respectively, in comparison with those of wild-type strain. Additionally, H2 production rate of KS0510 mutant was 4.8-fold higher than that of wild-type strain. The PTN0510 was identified to be much stronger than the well-known two strong promoters, gdh and slp promoters from Thermococcus strains, through RT-qPCR and western blotting analyses and kinetics of H2 production. In this study, we demonstrated that the RNA-seq approach is a good strategy to mine a novel strong promoter of use to a Thermococcus strain when developed as a biotechnologically promising strain to produce v | - |
dc.description.uri | 2 | - |
dc.language | English | - |
dc.publisher | 한국미생물학회 | - |
dc.relation.isPartOf | 한국미생물학회 국제학술대회 | - |
dc.title | Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon | - |
dc.type | Conference | - |
dc.citation.conferencePlace | KO | - |
dc.citation.endPage | 190 | - |
dc.citation.startPage | 190 | - |
dc.citation.title | 한국미생물학회 국제학술대회 | - |
dc.contributor.alternativeName | 이성혁 | - |
dc.contributor.alternativeName | 김민식 | - |
dc.contributor.alternativeName | 정해창 | - |
dc.contributor.alternativeName | 이정현 | - |
dc.contributor.alternativeName | 이현숙 | - |
dc.contributor.alternativeName | 강성균 | - |
dc.identifier.bibliographicCitation | 한국미생물학회 국제학술대회, pp.190 | - |
dc.description.journalClass | 2 | - |