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pcDNA4c/His-Xpress-HA-hBIG1(E793K)
(Plasmid #79433)

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Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 79433 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA4c
  • Backbone manufacturer
    ThermoFisher Scientific
  • Vector type
    Mammalian Expression
  • Selectable markers
    Zeocin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Insert is somewhat toxic to E. coli. Requires long incubation time to see colonies after transformation and only small colonies should be chosen for use. Fresh colonies (direct from incubator; not stored at 4C) must be used for growth in liquid culture. See depositor’s comments for additional info
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    BIG1
  • Species
    H. sapiens (human)
  • Mutation
    changed Glutamic acid 793 to Lysine
  • Entrez Gene
    ARFGEF1 (a.k.a. ARFGEP1, BIG1, P200)
  • Tags / Fusion Proteins
    • HA (N terminal on insert)
    • His6 (N terminal on backbone)
    • Xpress (N terminal on backbone)

Terms and Licenses

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

Depositor Comments

It is tricky to amplify the plasmid. After transformation of the plasmid into E. coli, not only the transformants grow very slowly (usually, it takes more than 20 h for the transformants to form visible colonies on ampicillin‐containing plates and colony size is smaller than usual one), but also their viability is drastically decreased during storage at 4C. Therefore, we routinely use fresh transformants for plasmid preparation. The plasmid yield decreases with the progress of the day after transformation. It is not recommended to make a pre-culture (~ 2ml) from a single colony for large scale culture. We strongly recommend that after transformation and recovery culture (for 1hr at 37C), bring all transformants to large scale culture (~ 200 ml) directly containing ampicillin.

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:

    pcDNA4c/His-Xpress-HA-hBIG1(E793K) was a gift from Kazuhisa Nakayama (Addgene plasmid # 79433 ; http://n2t.net/addgene:79433 ; RRID:Addgene_79433)
  • For your References section:

    BIG2, a guanine nucleotide exchange factor for ADP-ribosylation factors: its localization to recycling endosomes and implication in the endosome integrity. Shin HW, Morinaga N, Noda M, Nakayama K. Mol Biol Cell. 2004 Dec;15(12):5283-94. Epub 2004 Sep 22. 10.1091/mbc.e04-05-0388 PubMed 15385626