FIG. 5.

Mapping of the 5′ end of mRNA initiated from the tnpA promoter. The primer extension product is indicated by the arrow. Lanes 1 to 6 present primer extension reactions carried out with total RNA prepared from P. putida PRS2000 (lanes 1 and 2), P. putida PaW85 (lanes 3 and 4), and E. coli HB101 (lanes 5 and 6) carrying tnpA promoter-containing plasmid p1332S/C (lanes 1, 3, and 5) or pEST1332 (lanes 2, 4, and 6) as a negative control. Lanes C, T, A, and G show DNA sequencing reactions of plasmid p1332S/C; 26 nt of this sequence is presented at the left, and the transcription start point of the tnpA gene is marked by a diamond. DNA originated from the right end of Tn4652 in p1332S/C is indicated by the vertical bold line, and the −10 region of the tnpA promoter is boxed.