 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Apr 28, 2015 |
| Title |
Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon |
| Organism |
Thermococcus onnurineus NA1 |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
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.
|
| |
|
| Overall design |
RNA expreesion profile of T. onnurineus NA1 genes in CO-containing media, in duplicate, using Illumina HiSeq 2500
|
| |
|
| Contributor(s) |
Lee SH, Kim M, Jung HC, Lee J, Lee J, Lee HS, Kang SG |
| Citation(s) |
25690310 |
| |
| Submission date |
Dec 10, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Min-Sik Kim |
| E-mail(s) |
kms0540@kiost.ac
|
| Organization name |
Korea Institute of Ocean Science and Technology
|
| Department |
Department of Marine Biotechnology
|
| Street address |
Haean-ro
|
| City |
Ansan-si |
| State/province |
Gyeonggi-do |
| ZIP/Postal code |
426-744 |
| Country |
South Korea |
| |
|
| Platforms (1) |
| GPL19533 |
Illumina HiSeq 2500 (Thermococcus onnurineus NA1) |
|
| Samples (2) |
|
| Relations |
| BioProject |
PRJNA269922 |
| SRA |
SRP051024 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE64036_RAW.tar |
30.0 Kb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
|
|
|
|
 |
