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pET-15b PAI-RBP
(Plasmid #45390)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 45390 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET15b
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5700
  • Total vector size (bp) 6900
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Use AG1 or BL21(DE3)pLysS E. coli for protein overexpression.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    PAI-RBP
  • Alt name
    DKFZp564M2423
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1170
  • GenBank ID
    AL080119
  • Tag / Fusion Protein
    • His (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Nde I (not destroyed)
  • 3′ cloning site BamH I (not destroyed)
  • 5′ sequencing primer 5'-TAATACGACTCACTATAGGG-3' Novagen
  • 3′ sequencing primer 5'-GCTAGTTATTGCTCAGCGG-3' Novagen
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Resource Center and Primary Database of the German Human Genome Project
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

Our lab isolated the protein based on it's ability to bind to a region of the PAI-1 (plasminogen activator inhibitor type 1) 3'UTR that we had determined to be involved in the cAMP regulation of mRNA stability. A search of the database revealed that the cDNA for the protein had been cloned by the German Genome Project and dubbed hypothetical protein. They kindly provided us with the clone, which we subcloned into pET-15b for expression of the protein.
Subsequently, we discovered isoforms of the protein with or without 6 and/or 15 amino acid insertions. These variants are now subcloned into pCELF.

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:

    pET-15b PAI-RBP was a gift from Thomas Gelehrter (Addgene plasmid # 45390 ; http://n2t.net/addgene:45390 ; RRID:Addgene_45390)
  • For your References section:

    Identification and cDNA cloning of a novel RNA-binding protein that interacts with the cyclic nucleotide-responsive sequence in the Type-1 plasminogen activator inhibitor mRNA. Heaton JH, Dlakic WM, Dlakic M, Gelehrter TD. J Biol Chem. 2001 Feb 2;276(5):3341-7. Epub 2000 Sep 22. 10.1074/jbc.M006538200 PubMed 11001948