pMSP1D1
(Plasmid #20061)

• Purpose: Bacterial expression of membrane scaffold protein (MSP1D1) for Nanodisc formation
• Depositing Lab: Stephen Sligar
• Publication: Denisov et al J Am Chem Soc. 2004 Mar 24. 126(11):3477-87.

Backbone

• Vector backbone: pET 28a
• Backbone size w/o insert (bp): 5300
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: MSP1D1
• Alt name: membrane scaffold protein
• Alt name: APOA1
• Alt name: PDB: 1AV1
• Species: synthetic gene
• Insert Size (bp): 640
• Mutation: deletion mutant (1-11) of MSP1; His-tag on N-terminus followed by spacer sequence and TEV protease site
• Tag / Fusion Protein:
  • 7-His tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nco I (not destroyed)
• 3′ cloning site: Hind III (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 term

Resource Information

• Articles Citing this Plasmid:
  • 55 References

Terms and Licenses

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

Depositor Comments

NOTE: This plasmid is supplied in DH5alpha. For expression, it must be prepped and retransformed into BL-21(DE3); high oxygenation level

This plasmid has been found to be somewhat unstable and prone to concatenation. Concatenation often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.