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Addgene

pPFO*
(Plasmid #120400)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 120400 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pRSET-B
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 2887
  • Total vector size (bp) 4417
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    BL21 (DE3) pLysS E. coli competent cells used for protein production. Growth at 37°C until O.D. of 0.6 to 0.8. Then, expression induced with 1 mM IPTG at 37°C for 4 h.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Mutant form of Perfringolysin O
  • Alt name
    PFO*
  • Species
    Clostridium Perfringens
  • Insert Size (bp)
    1530
  • Mutation
    Amino acids 1-27 are deleted, starts at amino acid 28. Additionally, has three mutations: A71T, C459A, Y181A.
  • GenBank ID
  • Tag / Fusion Protein
    • His6 tag (N terminal on insert)

Cloning Information

  • Cloning method Unknown
  • 5′ sequencing primer TAA TAC GAC TCA CTA TAG GG
  • 3′ sequencing primer GCT AGT TAT TGC TCA GCG G
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Parent plasmid was obtained from Arthur Johnson, Texas A&M University. Site-directed mutagenesis was used to introduce 3 mutations.

Terms and Licenses

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

    pPFO* was a gift from Arun Radhakrishnan (Addgene plasmid # 120400 ; http://n2t.net/addgene:120400 ; RRID:Addgene_120400)
  • For your References section:

    Use of mutant 125I-perfringolysin O to probe transport and organization of cholesterol in membranes of animal cells. Das A, Goldstein JL, Anderson DD, Brown MS, Radhakrishnan A. Proc Natl Acad Sci U S A. 2013 Jun 25;110(26):10580-5. doi: 10.1073/pnas.1309273110. Epub 2013 Jun 10. 10.1073/pnas.1309273110 PubMed 23754385