Last data update: 24 January 2024 16:39 CET
Plasmid name: pENTRYL1L2-GAS1-Grx1-roGFP2 (LMBP 9706)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Caenorhabditis elegans general anaesthetic sensitivity abnormal cDNA (gas-1) Glutaredoxin cDNA (GLRX, GRX1) Aequorea victoria green fluorescent protein DNA (GFP); reduction-oxidation sensitive variant (roGFP), cytosol variant (roGFP2) |
| Promoter: | - |
| Ribosome binding site: |
- |
| Terminator: | Escherichia coli rrnB operon T1 terminator Escherichia coli rrnB operon T2 terminator |
| Selection marker: | Neomycin (neo; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pDONR201 |
| Further information: | This Gateway Entry vector was created by cloning the gas-1-GRX1-roGFP2 cassette into the pDONR201 vector. The reduction-oxidation sensitive green fluorescent protein (roGFP) is a redox sensitive biosensor. Two cysteines were introduced into the beta barrel structure of the GFP. The oxidation state (reduced dithiol or the oxidized disulfide) of the engineered thiols determines the fluorescence properties of the sensor. RoGFP2 preferentially interacts with glutaredoxins and therefore reports the cellular glutathione redox potential. The specificity of roGFP2 for glutathione is further increased by linking it to GRX1. By expressing the GRX1-roGFP fusion sensors in the organism of interest and/or targeting the protein to a cellular compartment, it is possible to measure the glutathione redox potential in a specific cellular compartment in real-time and therefore provides major advantages compared to other invasive static methods e.g. HPLC. The nucleotide sequence of the C. elegans insert can be retrieved from the Genbank accession number NC_003284.9. Other name of the plasmid is pENTRYL1L2GAS1TRANSGrx1-roGFP2. |
| EMBL Accession number: | NC_003284.9, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 09/12/2015 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by C. Vlaeminck(1) and Prof. Dr B. Braeckman(1). (1) Research Group for aging physiology and molecular evolution, Department of Biology, Ghent University, Ghent |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + kanamycin (50 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pENTRYL1L2-GAS1-Grx1-roGFP2 (LMBP 9706) is available at BCCM/GeneCorner. This plasmid was deposited by C. Vlaeminck and Prof. Dr B. Braeckman. |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.