Saved in:
| Main Authors: | , |
|---|---|
| Format: | Dataset Open Access |
| Language: | en |
| Published: |
PANGAEA
2021
|
| Subjects: | |
| Online Access: | https://doi.org/10.1594/PANGAEA.937559 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1867168213393997824 |
|---|---|
| author | Böhnke-Brandt, Stefanie Perner, Mirjam |
| author_facet | Böhnke-Brandt, Stefanie Perner, Mirjam |
| collection | Datos científicos de ciencias marinas y ambientales |
| contents | Here we used the first solely activity-based approach for identifying RubisCO active fosmid clones from a metagenomic library. Hydrothermal vent fluids derived from the interface zone between hot fluids emanating from Drachenschlund and ambient cold seawater at 8°18'S/13°30'W served as initial sample for library construction. Among 1056 screened fosmid clones 12 exhibited RubisCO activity. The metagenomic fragments of all twelve clones resembled genes from Hydrogenovibrio crunogenus. One of these clones was further analyzed. It contained a 35.2 kb metagenomic insert carrying the RubisCO gene cluster and flanking DNA regions. Knockouts of 12 genes and 2 intergenic regions on this metagenomic fragment demonstrated that the RubisCO activity was significantly impaired and was attributed to deletions in genes encoding putative transcriptional regulators and those believed to be vital for RubisCO activation. Our new technique revealed a novel link between a poorly-characterized gene and RubisCO activity. |
| format | Dataset Open Access |
| id | pangaea_https___doi_org_10_1594_PANGAEA_937559 |
| institution | PANGAEA |
| language | en |
| publishDate | 2021 |
| publisher | PANGAEA |
| record_format | pangaea |
| spellingShingle | Specific RubisCO activities of yet uncultured Proteobacteria depending on flanking genes Böhnke-Brandt, Stefanie Perner, Mirjam activity-based RubisCO screen; autotrophic CO2-fixation; Deleted open-reading frame; DERIDGE; From Mantle to Ocean: Energy-, Material- and Life-cycles at Spreading Axes; functional metagenomic; Gaussian propagation of error; High Performance Liquid Chromatography (HPLC); HPLC; hydrothermal deep sea vent; Identification; Insertion position; Insert size; Linkage between the distribution and biochemical properties of RuBisCO and CODH enzymes and abiotic properties in thermally and chemically distinct deep-sea hydrothermal vent systems; M78/2; M78/2_314; MARSUED5; Meteor (1986); Remote operated vehicle; ROV; RuBisCO; Sanger Sequencing modified after Sanger et al. (1977); PacBio Sequencing according to Rhoads and Au (2015); Sequence editing, BioEdit; Specific ribulose-1,5-bisphosphate carboxylase/oxygenase activity; Specific ribulose-1,5-bisphosphate carboxylase/oxygenase activity, standard deviation; Thiomicrospira crunogena; Total length of deleted open-reading frame Here we used the first solely activity-based approach for identifying RubisCO active fosmid clones from a metagenomic library. Hydrothermal vent fluids derived from the interface zone between hot fluids emanating from Drachenschlund and ambient cold seawater at 8°18'S/13°30'W served as initial sample for library construction. Among 1056 screened fosmid clones 12 exhibited RubisCO activity. The metagenomic fragments of all twelve clones resembled genes from Hydrogenovibrio crunogenus. One of these clones was further analyzed. It contained a 35.2 kb metagenomic insert carrying the RubisCO gene cluster and flanking DNA regions. Knockouts of 12 genes and 2 intergenic regions on this metagenomic fragment demonstrated that the RubisCO activity was significantly impaired and was attributed to deletions in genes encoding putative transcriptional regulators and those believed to be vital for RubisCO activation. Our new technique revealed a novel link between a poorly-characterized gene and RubisCO activity. |
| title | Specific RubisCO activities of yet uncultured Proteobacteria depending on flanking genes |
| topic | activity-based RubisCO screen; autotrophic CO2-fixation; Deleted open-reading frame; DERIDGE; From Mantle to Ocean: Energy-, Material- and Life-cycles at Spreading Axes; functional metagenomic; Gaussian propagation of error; High Performance Liquid Chromatography (HPLC); HPLC; hydrothermal deep sea vent; Identification; Insertion position; Insert size; Linkage between the distribution and biochemical properties of RuBisCO and CODH enzymes and abiotic properties in thermally and chemically distinct deep-sea hydrothermal vent systems; M78/2; M78/2_314; MARSUED5; Meteor (1986); Remote operated vehicle; ROV; RuBisCO; Sanger Sequencing modified after Sanger et al. (1977); PacBio Sequencing according to Rhoads and Au (2015); Sequence editing, BioEdit; Specific ribulose-1,5-bisphosphate carboxylase/oxygenase activity; Specific ribulose-1,5-bisphosphate carboxylase/oxygenase activity, standard deviation; Thiomicrospira crunogena; Total length of deleted open-reading frame |
| url | https://doi.org/10.1594/PANGAEA.937559 |