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pQE80L::smRatioGFP
(Plasmid #258589)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 258589 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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Backbone

  • Vector backbone
    pQE80L-Kan
  • Backbone manufacturer
    Qiagen
  • Backbone size w/o insert (bp) 4541
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Depositing lab recommends E. coli BL21(DE3)pLysS for expression.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    soluble modified ratiometric pHluorin
  • Species
    A. victoria
  • Insert Size (bp)
    739
  • GenBank ID
    AF058694
  • Promoter T5
  • Tag / Fusion Protein
    • 6xHis (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (unknown if destroyed)
  • 3′ cloning site PstI (unknown if destroyed)
  • 5′ sequencing primer GTATCACGAGGCCCTTTCGTCT
  • 3′ sequencing primer CATTACTGGATCTATCAACAGGAG
  • (Common Sequencing Primers)

Terms and Licenses

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

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

pQE80L::smRatioGFP A vector for bacterial expression, purification and in vitro analysis of the ratiometric At-pHluorin protein (Gao et al. 2004; DOI: 10.1104/pp.103.032508) and in particular for spectroscopic comparison with PtGFP (Schulte et al, doi.org/10.1186/1746-4811-2-7) The gene was originally designed for expression in Arabidopsis thaliana (for details see Addgene #240450 to #240454). The smRatioGFP gene is in frame with the preceeding sequence coding for a 6xHis-Tag. Sources for ratiometric GFP (ratiometric pHluorin): https://www.nature.com/articles/BF28190; soluble modified for expression in plants: https://pubmed.ncbi.nlm.nih.gov/9122158/ and https://pubmed.ncbi.nlm.nih.gov/9484447/.

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:

    pQE80L::smRatioGFP was a gift from Christoph Plieth (Addgene plasmid # 258589 ; http://n2t.net/addgene:258589 ; RRID:Addgene_258589)

  • 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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