The influence of ribosome-binding-site elements on translational efficiency in Bacillus subtilis and Escherichia coli in vivo

1992 ◽  
Vol 6 (9) ◽  
pp. 1105-1114 ◽  
Author(s):  
Robert Luis Vellanoweth ◽  
Jesse C. Rabinowitz
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Binding Site ◽  
Ribosome Binding Site ◽  
Translational Efficiency ◽  
Ribosome Binding

scholarly journals Identification of an Intragenic Ribosome Binding Site That Affects Expression of the uncB Gene of the Escherichia coli Proton-Translocating ATPase (unc) Operon

1998 ◽  
Vol 180 (15) ◽  
pp. 3940-3945 ◽  
Author(s):  
Sharlene R. Matten ◽  
Thomas D. Schneider ◽  
Steven Ringquist ◽  
William S. A. Brusilow
Keyword(s):  
Escherichia Coli ◽  
Binding Site ◽  
Fusion Gene ◽  
Ribosome Binding Site ◽  
Proton Channel ◽  
Translational Initiation ◽  
Reading Frame ◽  
Ribosome Binding ◽  
Codon Reading

ABSTRACT The uncB gene codes for the a subunit of the Fo proton channel sector of the Escherichia coli F1 Fo ATPase. Control of expression of uncB appears to be exerted at some step after translational initiation. Sequence analysis by the perceptron matrices (G. D. Stormo, T. D. Schneider, L. Gold, and A. Ehrenfeucht, Nucleic Acids Res. 10:2997–3011, 1982) identified a potential ribosome binding site within the uncB reading frame preceding a five-codon reading frame which is shifted one base relative to theuncB reading frame. Elimination of this binding site by mutagenesis resulted in a four- to fivefold increase in expression of an uncB′-′lacZ fusion gene containing most ofuncB. Primer extension inhibition (toeprint) analysis to measure ribosome binding demonstrated that ribosomes could form an initiation complex at this alternative start site. Two fusions oflacZ to the alternative reading frame demonstrated that this site is recognized by ribosomes in vivo. The results suggest that expression of uncB is reduced by translational frameshifting and/or a translational false start at this site within the uncB reading frame.

scholarly journals A Novel Bacillus subtilis Gene,antE, Temporally Regulated and Convergent to and Overlapping dnaE

1999 ◽  
Vol 181 (1) ◽  
pp. 353-356 ◽  
Author(s):  
Lin-Fa Wang ◽  
Sung-Soo Park ◽  
Roy H. Doi
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Binding Site ◽  
Ribosome Binding Site ◽  
Small Protein ◽  
Ribosome Binding ◽  
Untranslated Sequence ◽  
Partial Homology ◽  
Subtilis Gene

ABSTRACT A Bacillus subtilis promoter, Px, that functions in a convergent manner with the sigA operon promoter P3 has been found in the sigA operon. Promoter Px is turned on at the same time as promoter P3 during early sporulation. The transcript from promoter Px codes for a small protein with partial homology to the OmpR protein from Escherichia coli and also carries an untranslated sequence at its 3′ end that is complementary to the 5′ end of the P3 transcript, which codes for the ribosome binding site ofdnaE. The gene controlled by Px has been calledantE. The expression of antE does not require ςB, ςE, or ςH. Px was transcribed in vitro by the ςA holoenzyme and is the seventh promoter to be recognized in the ςA operon. A possible role for the antE gene during early sporulation is proposed.

pGR71 plasmid promotor sequence temporally regulated in Bacillus subtilis

1998 ◽  
Vol 44 (12) ◽  
pp. 1186-1192
Author(s):  
Guy Daxhelet ◽  
Philippe Gilot ◽  
Etienne Nyssen ◽  
Philippe Hoet
Keyword(s):  
Bacillus Subtilis ◽  
Binding Site ◽  
Transcription Initiation ◽  
Promoter Sequence ◽  
Initiation Site ◽  
Chloramphenicol Acetyltransferase ◽  
Ribosome Binding Site ◽  
Chloramphenicol Resistance ◽  
E Coli ◽  
Ribosome Binding

pGR71, a composite of plasmids pUB110 and pBR322, replicates in Escherichia coli and in Bacillus subtilis. It carries the chloramphenicol resistance gene (cat) from Tn9, which is not transcribed in either host by lack of a promoter. The cat gene is preceded by a Shine-Dalgarno sequence functional in E. coli but not in B. subtilis. Deleted pGR71 plasmids were obtained in B. subtilis when cloning foreign viral DNA upstream of this cat sequence, as well as by BAL31 exonuclease deletions extending upstream from the cat into the pUB110 moiety. These mutant plasmids expressed chloramphenicol acetyltransferase (CAT), conferring on B. subtilis resistance to high chloramphenicol concentrations. CAT expression peaked at the early postexponential phase of B. subtilis growth. The transcription initiation site of cat, determined by primer extension, was located downstream of a putative promoter sequence within the pUB110 moiety. N-terminal amino acid sequencing showed that native CAT was produced by these mutant plasmids. The cat ribosome-binding site, functional in E. coli, was repositioned within the pUB110 moiety and had consequently an extended homology with B. subtilis 16S rRNA, explaining the production of native enzyme.Key words: chloramphenicol acetyltransferase, Bacillus subtilis, postexponential gene expression, plasmid pUB110, ribosome-binding site, transcriptional promoter.

scholarly journals Efficient engineering of chromosomal ribosome binding site libraries in mismatch repair proficient Escherichia coli

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Sabine Oesterle ◽  
Daniel Gerngross ◽  
Steven Schmitt ◽  
Tania Michelle Roberts ◽  
Sven Panke
Keyword(s):  
Escherichia Coli ◽  
Binding Site ◽  
Mismatch Repair ◽  
Ribosome Binding Site ◽  
Ribosome Binding

scholarly journals Regulated Expression of the Escherichia coli dam Gene

2003 ◽  
Vol 185 (16) ◽  
pp. 5012-5014 ◽  
Author(s):  
Melissa A. Calmann ◽  
M. G. Marinus
Keyword(s):  
Escherichia Coli ◽  
Binding Site ◽  
Ribosome Binding Site ◽  
Lon Protease ◽  
Ribosome Binding ◽  
Regulated Expression

ABSTRACT Regulated expression of the Escherichia coli dam gene has been achieved with the araBAD promoter lacking a ribosome binding site. Cultures of dam mutants containing plasmid pMQ430 show no detectable methylation in the absence of arabinose and complete methylation in its presence. Dam methyltransferase is a substrate for the Lon protease.

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