Fig. 6. Determination of transcriptional start sites of the Manduca sexta
V-ATPase genes mvB, mvG and mvd. The start sites were
determined by RNase protection assays using 973 bp each of the 5'
upstream regions as a template for in vitro transcription of
32P-labelled ssRNA probes. After hybridization with poly(A) RNA,
single-stranded RNA was digested with RNase A and T1. The remaining
double-stranded RNA was separated on a denaturing polyacrylamide gel.
Visualization of radioactively labelled bands was performed by exposing the
gels to X-ray film. Nucleotide positions of identified transcriptional
initiation sites that are similar to the consensus sequence KCABHYBY
(Bucher, 1990) are shown on the
right side of the figure.
