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pRK1035
(Plasmid #8832)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 8832 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pMal-C2
  • Backbone manufacturer
    New England Biolabs
  • Backbone size w/o insert (bp) 6700
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    BL21(DE3)
  • Growth instructions
    Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TVMV protease.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    TVMV protease
  • Alt name
    tobacco vein mottling virus protease
  • Species
    tobacco vein mottling virus
  • Insert Size (bp)
    750
  • Mutation
    synthetic gene with codons optimized for E. coli
  • Tags / Fusion Proteins
    • MBP (cleaved) (N terminal on insert)
    • His (N terminal on insert)

Cloning Information

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
  • Article Citing this Plasmid

Depositor Comments

pRK1035 overproduces the catalytic domain of tobbaco vein mottling virus (TVMV) protease in the form of an MBP fusion protein that cleaves itself in vivo to yield a TVMV protease catalytic domain with an N-terminal His-tag. The specificity of TVMV protease is distinct from that of TEV protease. The canonical recognition site for TVMV protease is ETVRFQS, with cleavage occurring between Q and S. Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TVMV protease. The TVMV protease open reading frame on pRK1035 was constructed from synthetic oligonucleotides and has been codon-optimized for expression in E. coli.

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:

    pRK1035 was a gift from David Waugh (Addgene plasmid # 8832 ; http://n2t.net/addgene:8832 ; RRID:Addgene_8832)
  • For your References section:

    Efficient site-specific processing of fusion proteins by tobacco vein mottling virus protease in vivo and in vitro. Nallamsetty S, Kapust RB, Tozser J, Cherry S, Tropea JE, Copeland TD, Waugh DS. Protein Expr Purif. 2004 Nov . 38(1):108-15. 10.1016/j.pep.2004.08.016 PubMed 15477088