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M26I DJ-1 TEV site pET15b
(Plasmid #60685)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 60685 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET15b
  • Backbone manufacturer
    Novagen
  • Total vector size (bp) 5708
  • Vector type
    Bacterial 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
    M26I DJ-1
  • Alt name
    PARK 7
  • Species
    H. sapiens (human)
  • Mutation
    M26I and TEV site was inserted (see comments below).
  • Entrez Gene
    PARK7 (a.k.a. DJ-1, DJ1, GATD2, HEL-S-67p)
  • Promoter T7
  • Tag / Fusion Protein
    • 6xHis (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer T7
  • 3′ sequencing primer T7term
    • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This construct contains a Tobacco etch virus (TEV) protease recognition sequence (ENLYFQS) after the Thrombin recognition site, and just before DJ-1. (Ex. LVPRGSHMENLYFQSASKRAL......) with the TEV site italicized. Internal NdeI site of human DJ-1 eliminated by silent mutation (198A->T).

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:

    M26I DJ-1 TEV site pET15b was a gift from Mark Wilson (Addgene plasmid # 60685 ; http://n2t.net/addgene:60685 ; RRID:Addgene_60685)

  • For your References section:

    Transient sampling of aggregation-prone conformations causes pathogenic instability of a parkinsonian mutant of DJ-1 at physiological temperature. Milkovic NM, Catazaro J, Lin J, Halouska S, Kizziah JL, Basiaga S, Cerny RL, Powers R, Wilson MA. Protein Sci. 2015 Oct;24(10):1671-85. doi: 10.1002/pro.2762. Epub 2015 Aug 17. 10.1002/pro.2762 PubMed 26234586
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