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pET28a-6xHis-TrxF
(Plasmid #207454)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 207454 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET28a
  • Backbone size w/o insert (bp) 5293
  • Total vector size (bp) 5827
  • Modifications to backbone
    None
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Thioredoxin F
  • Alt name
    TrxF
  • Species
    Synthetic; Mesembryanthemum crystallinum
  • Insert Size (bp)
    534
  • Mutation
    Codon optimized for E coli expression
  • Promoter T7
  • Tag / Fusion Protein
    • 6xHis (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NheI (not destroyed)
  • 3′ cloning site xhoI (not destroyed)
  • 5′ sequencing primer T7 promoter
  • 3′ sequencing primer T7 terminator
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

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

Depositor Comments

Expression protocol: Transformed BL21-DE3 was grown to OD600 0.6-0.8, induced with 1mM IPTG and incubated at 12oC overnight. TrxF was purified by Ni-NTA affinity chromatography in buffer containing 50mM HEPES pH 8.0, 0.5M NaCl, 5% glycerol, 10mM β-mercaptoethanol and 5-300mM imidazole, then dialyzed into the same buffer without imidazole and stored in aliquots at -80oC. TrxF activity was verified using the insulin reduction assay (Holmgren, 1979).

Please visit https://www.biorxiv.org/content/10.1101/2022.08.24.505184v1for bioRxiv preprint.

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:

    pET28a-6xHis-TrxF was a gift from Joseph Jez & Sarah Stainbrook (Addgene plasmid # 207454 ; http://n2t.net/addgene:207454 ; RRID:Addgene_207454)

  • For your References section:

    C4 Grasses Employ Various Strategies to Acclimate Rubisco Activase to Heat Stress. Stainbrook SC, Aubuchon LN, Chen A, Johnson E, Si A, Walton L, Ahrendt A, Jez J. bioRxiv 2022.08.24.505184 10.1101/2022.08.24.505184
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