PGP382

pGP382: click to enlarge

pGP382 cloning region: click to enlarge

Application:

The vector was constructed in the Stülke lab and it is suitable for constitutive overexpression of C-terminally Strep-tagged proteins in B. subtilis. The plasmid confers resistance to ampicillin and erythromycin in E. coli and B. subtilis, respectively. pGP382 can be used for the SPINE method. pGP380 is similar to the vector pGP382 which is derived from vectors pBQ200 and pHT315. A Shine-Dalgarno sequence has to be fused to the open reading frame during PCR.

Sequencing primers:

• M13_puc_for: 5‘-GTAAAACGACGGCCAGTG-3‘
• M13_puc_rev: 5‘-GGAAACAGCTATGACCATG-3‘
• FX125 (rev; primes -114bp of MCS): 5‘-GGCTCGTATGTTGTGTGG-3‘
• JN54 (fwd; primes +63bp of MCS): 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘

Christina Herzberg, Lope Andrés Flórez Weidinger, Bastian Dörrbecker, Sebastian Hübner, Jörg Stülke, Fabian M Commichau
SPINE: a method for the rapid detection and analysis of protein-protein interactions in vivo.
Proteomics: 2007, 7(22);4032-5
[PubMed:17994626] [WorldCat.org] [DOI] (P p)