pBRα-β flap (831-1057)
PurposeExpression of alpha NTD fused via three alanines to E. coli RNAP subunit beta, aa 831-1057 (β-flap). Can be cut with Not1 and BamHI to replace β-flap with a moiety of interest.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53734||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector typeBacterial Expression
Growth in Bacteria
Growth Strain(s)NEB5alpha F'LacIQ
Growth instructionsWe recommend using a lacIQ strain, such as NEB5alpha F'LacIQ, in order to ensure that the expression of potentially toxic fusion proteins is kept repressed.
Gene/Insert namealpha NTD-Beta flap fusion
Alt namerpoA-rpoB fusion
- Promoter lpp/lacUV5 (tandem promoter)
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer pBRa_F (approx 150bp upstream of NotI site) 5’ gaacagcgtaccgacctggac 3’ (Common Sequencing Primers)
Note-When cloning into this plasmid, the 5' end of the insert must provide an extra nucleotide to maintain the reading frame. Additionally, a stop codon must be inserted 5' to the BamHI site. Please see supplemental document for more details.
This plasmid is used in conjuction with the other Ann Hochschild Bacterial 2-hybrid system plasmids (Addgene plasmids 53730, 53731, 53732, 53733, and 53734) and must be used in the reporter strain, FW102 OL2–62 (Addgene item 53735). Please see the associated article for links to all of these plasmids:
The following articles can provide more information on using the bacterial 2-hybrid system:
Dove, et al. 1997 PMID 9121589
Dove and Hochschild 1998 PMID 9499408
Deaconescu, et al. 2006 PMID 16469698
Deighan, et al. 2008 PMID 18832144
Kuznedelov, et al. 2002 PMID 11823642
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:pBRα-β flap (831-1057) was a gift from Ann Hochschild (Addgene plasmid # 53734 ; http://n2t.net/addgene:53734 ; RRID:Addgene_53734)
For your References section:Activation of prokaryotic transcription through arbitrary protein-protein contacts. Dove SL, Joung JK, Hochschild A. Nature. 1997 Apr 10;386(6625):627-30. 10.1038/386627a0 PubMed 9121589