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. 2009 Dec 29;38(6):1950–1963. doi: 10.1093/nar/gkp1190

Figure 2.

Figure 2.

Characterization of promoter recruitment and of run-off transcripts of ΔH RNAP. (A) ΔH RNAP forms stable preinitiation complexes. EMSAs with a probe containing the Pyrococcus gdh promoter (10) were conducted in the presence and absence of TBP, TFB and reconstituted wt or ΔH RNAP as indicated on top of the lanes. The position of the TBP–TFB and TBP–TFB–RNAP complexes are indicated. (B) Eucaryotic Rpb5 can functionally replace RpoH in an archaeal RNAP. The synthesis of a 145 nt run-off transcript from the Pyrococcus gdh promoter was analyzed in standard multiple round transcription assays (‘Materials and Methods’ section) and RNA products were analyzed on 6% denaturing PA gels. Lane 4 shows transcription products synthesized by ΔH RNAP reconstituted with Rpb5. In lanes 2 and 3 subunit H or Rpb 5 were added to transcription reactions. The diagram below the gel shows the mean value of the transcriptional activity for each lane. The quantification was done with the Aida image analyzer software version 3.28.