Skip to main content
. 2005 Sep 12;33(16):e139. doi: 10.1093/nar/gni140

Figure 3.

Figure 3

T7RNAP-driven transcription assay by in organello RNA synthesis after electroporation into isolated mouse T7RNAP-mitochondria. (A) Structure of pUC-T7CAT, a control plasmid containing the prokaryotic CAT gene and T7 promoter (PT7). The CAT gene is transcribed from the T7 promoter when this plasmid is introduced into transcriptionally active T7RNAP-mitochondria. (B) RT–PCR analysis of the CAT transcripts in electroporated mouse T7RNAP-mitochondria. Plasmid pUC-T7CAT was electroporated into the isolated mouse T7RNAP-mitochondria and in organello RNA syntheses of these electroporated mitochondria were performed in incubation buffer for 2 h at 37°C. After isolating total RNA from the mitochondria, RT–PCR was carried out using CAT-specific primers CmR-F and CmR-R (see Materials and Methods). Expected band size of the RT–PCR CAT products was 470 bp. Lane M, 1 kb plus DNA ladder (Life Technologies); lanes 1 and 2, no electroporation control with plasmid; lanes 3 and 4, 12 kV/cm electroporation with plasmid. Control lanes in which reverse transcriptase (RT) was omitted are indicated below by minus signs.