pART7::PtGFP:PX
(Plasmid
#258580)
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PurposeTransient expression of the indicator protein PtGFP for monitoring the pH in the peroxisomes of plant cells.
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Depositing Lab
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 258580 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
Backbone
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Vector backbonepART7
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Backbone manufacturerAndrew Gleave (Addgene #247985)
- Backbone size w/o insert (bp) 5042
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Vector typeBacterial Expression, Plant Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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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)763
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GenBank IDAY015995
- Promoter CaMV 35S (GenBank AJ007625) without artificial 40bp insert; drives expression plants
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Tag
/ Fusion Protein
- SRL tripeptide (peroxisome targeting sequence) (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site XhoI (unknown if destroyed)
- 3′ cloning site HindIII (unknown if destroyed)
- 5′ sequencing primer CAACCACGTCTTCAAAGC
- 3′ sequencing primer GGATCTGAGCTACACATGC
- (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
pART7::PtGFP:PX is similar to pART7::PtGFP with the difference that the PtGFP gene is extended at its 3'-end by nine bases coding for the tripeptide 'SRL'. This tripeptide belongs to a class of C-terminal signal sequences which direct expressed proteins to the peroxisomes in plants (Reumann 2004; doi.org/10.1104/pp.103.035584) 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:
pART7::PtGFP:PX was a gift from Christoph Plieth (Addgene plasmid # 258580 ; http://n2t.net/addgene:258580 ; RRID:Addgene_258580) -
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