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. 2007 Jun 29;189(17):6437–6446. doi: 10.1128/JB.00421-07

FIG. 6.

FIG. 6.

Two-hybrid analysis of the interaction between MamJ and MamK and MamJ and MamJ. (A) Growth of reporter E. coli spotted onto selective and nonselective screening medium after cotransformation with different bait (derivative of pBT) and prey (derivative of pTRG) expression vectors. Colony growth on nonselective screening medium verifies that cotransformation was successful, whereas on selective screening medium, colonies can grow only in the case of an interaction between bait and prey fusion proteins. (B) Overview of analyzed MamJ sequence regions (prey) for two-hybrid interaction with MamK, MamJ, and the N-terminal part of MamJ (bait). Growth on both selective and nonselective screening medium is shown by +; − indicates growth on nonselective screening medium only. n.d., not determined. (C) Immunodetection of prey fusion proteins by an anti-MamJ peptide antibody in protein crude extracts of the BacterioMatch reporter E. coli resolved by SDS-PAGE. RNAP fusions with J295-334, J330-368, and J361-399 lack the epitope recognized by the antibody.