pMBP-MS2
(Plasmid #65104)

• Purpose: can be used to isolate RNA binding complexes
• Depositing Lab: Josep Vilardell
• Publication: Macias et al Mol Cell. 2008 Jun 20;30(6):732-42. doi: 10.1016/j.molcel.2008.05.002.

Backbone

• Vector backbone: pMal-c
• Backbone manufacturer: NEB
• Total vector size (bp): 6442
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL-2 Blue
• Growth instructions: Cannot be grown in DH5 alpha. Has to be grown in a strain that requires IPTG induction of the promoter, as E. coli don't like it if it's constitutively expressed. TB1 strain is also good.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: MS2
• Species: Synthetic
• Mutation: double mutation (V75Q and A81G) that prevents oligomerization
• Promoter: tac
• Tag / Fusion Protein:
  • MBP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: StuI (destroyed during cloning)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13pUC-Rev or MBP-F
• 3′ sequencing primer: M13pUC-Fwd

Resource Information

• Supplemental Documents:
  • pMBP-MS2 seq
• Articles Citing this Plasmid:
  • 7 References

Terms and Licenses

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

Depositor Comments

The following reference describes how to isolate RNA binding complexes using this construct:
Jurica and Moore, Methods. 2002 Nov;28(3):336-45. https://www.ncbi.nlm.nih.gov/pubmed/12431437