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pART7::COX:PtGFP
(Plasmid #258581)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 258581 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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Backbone

  • Vector backbone
    pART7
  • Backbone manufacturer
    Andrew Gleave (Addgene #247985)
  • Backbone size w/o insert (bp) 5042
  • Vector type
    Bacterial Expression, Plant Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    PtGFP
  • Species
    S. cerevisiae (budding yeast); Ptilosarcus sp. CSG-2001
  • Insert Size (bp)
    927
  • GenBank ID
    AY015995, X01418
  • Promoter CaMV 35S (GenBank AJ007625) without artificial 40bp insert;144nt TEV enhancer (GenBank GU734657)
  • Tag / Fusion Protein
    • COX-IV signal sequence (N 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

  • A portion of this plasmid was derived from a plasmid made by
    Dr. Bruce Bryan, Prolume.com

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

pART7::COX:PtGFP is for targeting PtGFP to the plant mitochondria matrix. The PtGFP gene is concatenated downstream of a sequence coding for the signal sequence (first 29 aminoacids) of the cytochrome oxidase subunit IV from yeast (COX-IV; Gene Bank X01418; Maarse et al. 1984, doi.org/10.1002/j.1460-2075.1984.tb02216.x; Akashi et al. 1998, doi.org/10.1016/S0014-5793(98)00717-0; Filipin et al. 2005, doi.org/10.1016/j.ceca.2004.08.002; Llopis et al. 1998, doi.org/10.1073/pnas.95.12.6803 ). A 5'UTR 144 nucleotide translation leader sequence from the tobacco etch virus (TEV; Gene Bank GU734657) was prepended for enhancement of expression (Carrington and Freed 1990 doi.org/10.1128/jvi.64.4.1590-1597.1990; Zeenko and Gallie 2005, DOI: 10.1074/jbc.M503576200; Peeters et al. 2000, doi.org/10.1007/s002390010044; Harper et al. 1998, doi.org/10.1074/jbc.273.2.1099) 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).

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pART7::COX:PtGFP was a gift from Christoph Plieth (Addgene plasmid # 258581 ; http://n2t.net/addgene:258581 ; RRID:Addgene_258581)

  • 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
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