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hE-cadherin-pcDNA3
(Plasmid #45769)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 45769 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5446
  • Total vector size (bp) 8550
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    hE-cadherin
  • Alt name
    E-cadherin
  • Alt name
    CDH-1
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    3100
  • GenBank ID
    NM_004360.4 NP_004351.1
  • Entrez Gene
    CDH1 (a.k.a. Arc-1, BCDS1, CD324, CDHE, ECAD, LCAM, UVO)
  • Promoter CMV

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site XbaI (not destroyed)
  • 5′ sequencing primer CMV-F; T7
  • 3′ sequencing primer BGH-rev; Sp6
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Human E-cadherin partial cDNA provided by David Rimm (Yale University, New Haven, CT)
  • Terms and Licenses
  • Articles Citing this Plasmid

Depositor Comments

Partial cDNA for human E-cadherin was provided by D. Rimm (Yale University, New Haven, CT) and subcloned into the pcDNA3 mammalian expression vector (Invitrogen), to generate Addgene plasmid 45772. Sequence analysis revealed that the 3′ end of the gene was missing after nucleotide 2644 (according to EMBL/GenBank/DDBJ under accession number L08599). This resulted in a truncation of the last 35 amino acids of the E-cadherin cytoplasmic domain and, as a result, did not contain the β-catenin binding region, as defined by Stappert and Kemler 1994. The COOH terminus of this truncation mutant (E-cadherin Δ β-catenin) ends at amino acid 844 (NH3-ASLSSD); the frameshift added a single histidine residue before a stop codon is introduced.

The full-length human E-cadherin was reengineered using RNA from human A431 cells and the RT-PCR method
(Primer A, 5′-TGACACCCGGGACAACGTTTATTA-3′, and Primer C, 5′-CTAGTCTAGACCCCTAGTGGTCCTCG-3′)
to generate a 425-bp fragment encoding the missing COOH-terminal residues. This fragment was sub-cloned into the truncated hEcad-Δ 35/pcDNA3 vector (Addgene plasmid 45772) to generate full-length hEcad/pcDNA3 (Addgene plasmid 45769).

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:

    hE-cadherin-pcDNA3 was a gift from Barry Gumbiner (Addgene plasmid # 45769 ; http://n2t.net/addgene:45769 ; RRID:Addgene_45769)
  • For your References section:

    E-cadherin suppresses cellular transformation by inhibiting beta-catenin signaling in an adhesion-independent manner. Gottardi CJ, Wong E, Gumbiner BM. J Cell Biol. 2001 May 28;153(5):1049-60. 10.1083/jcb.153.5.1049 PubMed 11381089