FIGURE 4.
Toeprinting of the lpp, ompA, and mazG mRNA.
A, toeprinting of the lpp mRNA in the presence of
YoeB(His)6. B, toeprinting of the ompA mRNA in
the presence of YoeB(His)6. C, toeprinting of the
mazG mRNA in the presence of YoeB(His)6. For each panel,
lane 1, without YoeB(His)6, 70 S ribosomes, and
; lane
2, 3.4 μm YoeB(His)6 alone; lane 3,
0.05 μm 70 S ribosomes and 1 μm
; lane
4, 0.05 μm 70 S ribosomes and 1 μm
with 3.4
μm YoeB(His)6; lane 5, 0.05 μm
70 S ribosomes and 1 μm
with 3.4
μm YoeB(His)6 and 13.6 μm glutathione
S-transferase-YefM. The sequence ladders shown at the
right-hand side in all cases were obtained using the same primers
used for toeprinting with their corresponding genes cloned in pCR 2.1-TOPO as
template. All mRNA sequences shown are complementary to the sequencing
ladders. The initiation codon, AUG, is indicated with an arrowhead.
TP(Y) is the band where toeprinting was stopped in the
presence of YoeB(His)6. FL, the full-length of the mRNA;
and TP(r), the toeprinting site due to normal ribosome
binding to mRNA in the absence of YoeB(His)6. The band at TP(S) is
likely to be formed due to a secondary structure of the mRNA.