10.1371/journal.pone.0105749.g008
Tomotaka Tanabe
Tomotaka
Tanabe
Ayaka Kato
Ayaka
Kato
Keiichi Shiuchi
Keiichi
Shiuchi
Katsushiro Miyamoto
Katsushiro
Miyamoto
Hiroshi Tsujibo
Hiroshi
Tsujibo
Jun Maki
Jun
Maki
Shigeo Yamamoto
Shigeo
Yamamoto
Tatsuya Funahashi
Tatsuya
Funahashi
<i>In vitro</i> translation of <i>peuA</i> mRNA.
Public Library of Science
2014
pH 8.0
pH 7.0
translation stage
PeuA
complementation experiments
Secondary structure prediction
Vibrio parahaemolyticus peuA Gene Encoding
silico analyses
Primer extension analysis
Vibrio parahaemolyticus
ferric uptake regulator
TonB 2 system genes
translation analysis
deletion mutants
peuRS deletion
novel gene regulation mechanism
enterobactin utilization
system operon
undescribed receptor
peuA expression
site
vpa
transcript
Translation initiation
Alternative Ferric Enterobactin Receptor
system PeuRS
2014-08-22 03:34:55
Figure
https://plos.figshare.com/articles/figure/_In_vitro_translation_of_peuA_mRNA_/1149419
<p>(A) <i>In vitro</i> translation analysis of the +1 and +39 <i>peuA</i> transcripts labeled with the FLAG tag. The +1-<i>peuA</i>′-<i>flag</i> RNA and +39-<i>peuA</i>′-<i>flag</i> RNA were first synthesized by <i>in vitro</i> transcription, as described in the MATERIALS AND METHODS, and a mixture containing either the +1-<i>peuA</i>′-<i>flag</i> RNA (30 pmol)/<i>fur</i>-<i>flag</i> RNA (3 pmol) or the +39-<i>peuA</i>′-<i>flag</i> RNA (30 pmol)/<i>fur</i>-<i>flag</i> RNA (3 pmol) as the template was subjected to <i>in vitro</i> translation. The FLAG-fused proteins translated were separated on 15% SDS-polyacrylamide gels, and were detected by western blotting using anti-FLAG IgG. (B) Confirmation of the presence of +1-<i>peuA</i>′-<i>flag</i> RNA and +39-<i>peuA</i>′-<i>flag</i> RNA in the reaction mixture for <i>in vitro</i> translation. These RNA fragments were detected in the reaction mixture by northern blotting using a DIG-labeled <i>peuA</i> probe.</p>