EF.delmFasL.CMV.GFP
(Plasmid #17620)

• Depositing Lab: Linzhao Cheng
• Publication: Yu et al Mol Ther. 2003 Jun . 7(6):827-38.

Backbone

• Vector backbone: EF.CMV.GFP
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: GFP

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: delFasL
• Alt name: Fas ligand
• Species: M. musculus (mouse)
• Insert Size (bp): 800
• Mutation: Noncleavable mouse delFasL . Deleted the exon 2 region, which contains the metalloproteinase cleavage site.
• Entrez Gene: Fasl (a.k.a. APT1LG1, CD178, CD95-L, CD95L, Fas-L, Faslg, Tnfsf6, Tnlg1a, gld)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: EF-1a forward

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The CMV promoter was inserted immediately upstream of GFP in the EF.GFP LV to make the intermediate LV, EF.V-CMV.GFP, with a unique EcoRV site available to insert a gene of interest under the control of the EF promoter. Noncleavable mouse delFasL cDNA was constructed using PCR cloning to delete the exon 2 region, which contains the metalloproteinase cleavage site. The delFasL gene was then cloned into the EcoRV site of EF.CMV.GFP to make the EF.delFasL-CMV.GFP LV.

Recombinant LVs were produced by transient transfection of the transducing vector into 293T cells with two packaging vectors: pMD.G, a plasmid expressing the VSV-G envelope gene, and pCMVDeltaR8.91, a plasmid expressing the HIV-1 gag/pol, tat, and rev genes.