4.5 Article

Improvement of bacterial two-hybrid vectors for detection of fusion proteins and transfer to pBAD-tandem affinity purification, calmodulin binding peptide, or 6-histidine tag vectors

Journal

PROTEOMICS
Volume 8, Issue 22, Pages 4768-4771

Publisher

WILEY
DOI: 10.1002/pmic.200800270

Keywords

Bacterial two-hybrid; Protein-protein interaction; Protein detection

Funding

  1. CNRS
  2. FRM fellowship

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The original vectors of the bacterial two-hybrid technique developed by Karimova et al. in 1998 did not enable detection of the recombinant proteins. Here, we propose two methods resolving this problem, either using new plasmids containing the Flag epitope, or using a trick to detect the T18 domain of adenylate cyclase. Furthermore, we describe a set of vectors for TAP, CBP or 6-histidine tagging that possess the same cloning site as our two-hybrid vectors.

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