Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon

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 valuable metabolites through a metabolic engineering. RNA expreesion profile of T. onnurineus NA1 genes in CO-containing media, in duplicate, using Illumina HiSeq 2500

ORGANISM(S): Thermococcus onnurineus NA1

SUBMITTER: Min-Sik Kim

PROVIDER: E-GEOD-64036 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon.

Lee Seong Hyuk SH Kim Min-Sik MS Jung Hae Chang HC Lee Jinwon J Lee Jung-Hyun JH Lee Hyun Sook HS Kang Sung Gyun SG

Applied microbiology and biotechnology 20150218 9

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 from 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 severa ...[more]

PMID: 25690310

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Dataset Information

Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon

Publications

Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon.

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