pQE80L::PtGFP
(Plasmid
#258588)
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PurposeExpression of the pH indicator PtGFP in E. coli.
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Depositing Lab
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 258588 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
Backbone
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Vector backbonepQE80L-Kan
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Backbone manufacturerQiagen
- Backbone size w/o insert (bp) 4541
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsDepositing lab recommends E. coli BL21(DE3)pLysS for expression.
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePtGFP
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SpeciesPtilosarcus sp. CSG-2001
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Insert Size (bp)723
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GenBank IDAY015995
- Promoter T5
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Tag
/ Fusion Protein
- 6xHis (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (unknown if destroyed)
- 3′ cloning site HindIII (unknown if destroyed)
- 5′ sequencing primer GTATCACGAGGCCCTTTCGTCT
- 3′ sequencing primer CATTACTGGATCTATCAACAGGAG
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byDr. Bruce Bryan, Prolume.com
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
pQE80L::PtGFP — For bacterial expression aiming at protein purification via Ni2+-agarose and in vitro analyses of PtGFP, the commercial vector pQE80l-Kan from Qiagen was employed https://www.qiagen.com/en-us/resources/download/protocols/cis-repressed-pqe-vector-maps-en. The PtGFP gene is in frame with the preceeding sequence coding for a 6xHis-Tag. PtGFP (GenBank AY015995) is a green fluorescent protein from the sea pen Ptilosarcus gurneyi which has been expressed in plants and used as a ratiometric pH indicator (Schulte et al. 2006 doi.org/10.1186/1746-4811-2-7, PubMed ID: 16600023). PtGFP is more photostable and less sensitive to acidic environments when compared with pHluorins (GenBank AF058694; GenBank AF058695).
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pQE80L::PtGFP was a gift from Christoph Plieth (Addgene plasmid # 258588 ; http://n2t.net/addgene:258588 ; RRID:Addgene_258588) -
For your References section:
A novel fluorescent pH probe for expression in plants. Schulte A, Lorenzen I, Bottcher M, Plieth C. Plant Methods. 2006 Apr 6;2:7. doi: 10.1186/1746-4811-2-7. 10.1186/1746-4811-2-7 PubMed 16600023