RGD-ACP-I27-CC
(Plasmid #85391)

• Purpose: MTFM sensor to measure integrin mechanical forces. Integrin binding domain fused to titan immunoglobulin domain (I27) with two cysteines (CC) for immobilization and ACP tag for fluorophore labeling
• Depositing Lab: Khalid Salaita
• Publication: Galior et al Nano Lett. 2016 Jan 13;16(1):341-8. doi: 10.1021/acs.nanolett.5b03888. Epub 2015 Dec 7.

Backbone

• Vector backbone: pET-22b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5493
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use BL21(DE3) strain for expression
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: RGD motif for integrin binding fused to acyl carrier protein (ACP) tag and titin immunoglobulin domain (I27)
• Alt name: KS24-ECC
• Promoter: T7
• Tag / Fusion Protein:
  • 6x His (C terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7-term

Resource Information

• Supplemental Documents:
  • 85391.gb

Terms and Licenses

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

Depositor Comments

Fibronectin binding motif (RGD) is TVYAVTGRGDSPASSAA.

ACP-tag is present at the N-terminus of I27 domain and was used to label the fusion protein with the Alexa488 fluorophore.

I27 domain is the 27th immunoglobulin domain of cardiac titin. Two cysteine residues were added at the C-terminus of I27 to allow immobilization onto a gold nanoparticle.

Please see the associated publication for material and methods information.