Table 1. Effect of incubation conditions on transcription of pCox II by electrotransformed mitochondria.

Complete (+ 1µg DNA)	++++
Complete (+ 10 µg DNA)	+++
–Amino acids	++++
7.5 mM KCl; 5 mM MgCl2	+
–Succinate, –ADP	–
–Succinate, –ADP, +ATP (0.15 mM)	–
0.15mM GTP	++
–GTP	–
–rNTP	–

Purified wheat embryo mitochondria were electroporated in the presence of pCox II plasmid. The RNA was extracted and total cDNA was synthesized with random primers. The target cox II transcript was revealed by PCR using primers 1 and 3. The complete reaction mixture contained 1 µg DNA, 330 mM mannitol, 90 mM KCl, 10 mM MgCl₂, 12 mM tricine pH 7.2, 5 mM KH₂PO₄, 1.2 mM EGTA, 1 mM GTP, 2 mM DTT, 2 mM ADP, 10 mM sodium succinate, 0.15 mM each of CTP and UTP and 25 µM amino acid mixture. Relative transcription efficiency was determined by ethidium bromide UV fluorescence after gel electrophoresis. The data were normalized using the signal obtained with the endogenous nad1 transcripts (primers 6 and 7) as internal standard.