pMDLg/pRRE
(Plasmid #12251)

Available to Academic and Nonprofits Only

Backbone

  • Vector backbone
    pMD
  • Backbone size w/o insert (bp) 8895
  • Vector type
    Mammalian Expression, Lentiviral ; Packaging

Growth in Bacteria

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

Sequence Information

Gene/Insert

  • Gene/Insert name
    HIV-1 GAG/POL

Cloning Information

Resource Information

Depositor Comments

This 3rd generation LV packaging plasmid includes gag, coding for the virion main structural proteins; pol, responsible for the retrovirus-specific enzymes; and RRE, a binding site for the Rev protein which facilitates export of the RNA from the nucleus. This plasmid does not include Rev. The third generation packaging system offers maximal biosafety but is more cumbersome, as it involves the transfection of four different plasmids in the producer cells.
Example of a 3rd generation packaging combo: pMDLg/pRRE + pRSV-Rev + pMD2.G.

Please note that most Trono lab lentivectors contain a wt 5'LTR and can be packaged only using the 2nd generation packaging system (as the wt 5'LTR requires TAT for activation).
If you wish to use the 3rd generation packaging system, you need to have a lentivector with a chimeric 5'LTR (e.g. CCL-, RRL-, etc) in which the HIV promoter is replaced with CMV or RSV, thus making it TAT-independent. The lentivectors carrying the chimeric 5'LTR can be packaged into either a 2nd or 3rd generation packaging system.

Please note that the full sequence for this plasmid is approximated and not fully verified. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussions on cloning strategies and protocols.

Please note that the ClaI site in this plasmid is blocked by Dam methylation. Do not use this enzyme for diagnostic digests.

How to cite this plasmid

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:

    pMDLg/pRRE was a gift from Didier Trono (Addgene plasmid # 12251)
  • For your References section:

    A third-generation lentivirus vector with a conditional packaging system. Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. J Virol. 1998 Nov . 72(11):8463-71. PubMed 9765382