CD40L-MIEG-hCD4 (hCD4-CD40L)
(Plasmid #40355)

• Depositing Lab: Alexander Dent
• Publication: Kusam et al Int J Cancer. 2009 Aug 15;125(4):977-81.

Backbone

• Vector backbone: MIEG-hCD4
• Backbone manufacturer: Addgene plasmid 40569
• Modifications to backbone: IRES EGFP of pMIEG replaced with human CD4 cDNA
• Vector type: Retroviral

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: CD40L
• Alt name: CD40-ligand
• Alt name: CD154
• Species: M. musculus (mouse)
• Entrez Gene: Cd40lg (a.k.a. CD154, CD40-L, Cd40l, HIGM1, IGM, IMD3, Ly-62, Ly62, T-BAM, TRAP, Tnfsf5, Tnlg8b, gp39)
• Tag / Fusion Protein:
  • IRES hCD4 (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: XhoI (unknown if destroyed)
• 5′ sequencing primer: pLXSN_5
• 3′ sequencing primer: IRES-R2 (gacggcaatatggtggaaa)

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

The CD40-ligand (CD40L)-expressing retroviral vector was constructed by PCR amplification of cDNA from anti-CD3 activated mouse T cells with the following primers for CD40-ligand: sense 5'-CTTTCAGTCAGCATGATAGAAACA-3' and antisense
5'-TCAGAGTTTGAGTAAGCCAAAAGA-3'; these primers were used to amplify the complete coding region of mouse CD40 ligand. The CD40-ligand gene was then reamplified with primers containing XhoI and NotI sites and then cloned via XhoI and NotI sites into a retroviral vector expressing human CD4.