Purposecan be used to isolate RNA binding complexes
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||65104||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Total vector size (bp) 6442
Vector typeBacterial Expression
Growth in Bacteria
Growth Strain(s)XL-2 Blue
Growth instructionsCannot 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.
Mutationdouble mutation (V75Q and A81G) that prevents oligomerization
- Promoter tac
/ Fusion Protein
- MBP (N terminal on insert)
- 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 (Common Sequencing Primers)
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
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pMBP-MS2 was a gift from Josep Vilardell (Addgene plasmid # 65104 ; http://n2t.net/addgene:65104 ; RRID:Addgene_65104)
For your References section:L30 binds the nascent RPL30 transcript to repress U2 snRNP recruitment. Macias S, Bragulat M, Tardiff DF, Vilardell J. Mol Cell. 2008 Jun 20;30(6):732-42. doi: 10.1016/j.molcel.2008.05.002. 10.1016/j.molcel.2008.05.002 PubMed 18570876