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pART7::16TP:PtGFP
(Plasmid #258584)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 258584 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
    Ptilosarcus sp. CSG-2001, Spinacia oleracea
  • Insert Size (bp)
    1002
  • GenBank ID
    X05512, AY015995
  • Promoter CaMV 35S (GenBank AJ007625) without artificial 40bp insert; drives expression plants
  • Tag / Fusion Protein
    • 16kDa-TP transit peptide (83 aa) from the Spinach photosynthetic oxygen-evolving complex (OEC) (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::16TP:PtGFP — A transit peptide, namely the 16kDa-TP (Gene Bank: X05512) from the Spinach photosynthetic oxygen-evolving complex (OEC) was employed to direct the expression of PtGFP to the thylakoid lumen of chloroplasts (Jansen et al. 1987, doi.org/10.1016/0014-5793(87)80696-8; Clausmeyer et al. 1993, doi.org/10.1016/S0021-9258(19)85183-9; Marques et al. 2003, doi.org/10.1007/s00438-003-0846-y) The 249 nucleotides of the 16 kDa TP coding for 83 AAs from the OEC were concatened with the PtGFP gene. 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::16TP:PtGFP was a gift from Christoph Plieth (Addgene plasmid # 258584 ; http://n2t.net/addgene:258584 ; RRID:Addgene_258584)

  • 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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