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 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.

The T7 phage gene 10 leader RNA, a ribosome-binding site that dramatically enhances the expression of foreign genes in Escherichia coli

Gene ◽  
1988 ◽  
Vol 73 (1) ◽  
pp. 227-235 ◽  
Author(s):  
Peter O. Olins ◽  
Catherine S. Devine ◽  
Shaukat H. Rangwala ◽  
Kamilla S. Kavka
Keyword(s):  
Escherichia Coli ◽  
Binding Site ◽  
Ribosome Binding Site ◽  
Phage Gene ◽  
Ribosome Binding ◽  
Foreign Genes ◽  
T7 Phage

scholarly journals The pseudoazurin gene from Thiosphaera pantotropha: analysis of upstream putative regulatory sequences and overexpression in Escherichia coli

1997 ◽  
Vol 321 (3) ◽  
pp. 699-705 ◽  
Author(s):  
Yun-Chung LEUNG ◽  
Christopher CHAN ◽  
John S. READER ◽  
Anthony C. WILLIS ◽  
Rob J. M. van SPANNING ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Binding Site ◽  
Paracoccus Denitrificans ◽  
Regulatory System ◽  
Ribosome Binding Site ◽  
Regulatory Sequences ◽  
Ribosome Binding ◽  
Thiosphaera Pantotropha

The pseudoazurin gene from Thiosphaera pantotropha has been cloned and sequenced. The deduced amino acid sequence showed that the protein contains an unusually alanine-rich signal peptide, 22 amino acid residues in length, which targets the protein to the periplasm. This pseudoazurin was expressed in large amounts in the periplasm of Escherichia coli when the gene with its native ribosome-binding site was placed downstream of the lac promoter. Removal of a putative hairpin-forming structure upstream of the ribosome-binding site increased the yield of the purified protein to ∼80 mg/l. The recombinant protein is indistinguishable from that purified from its natural host. A primer extension study indicated that the pseudoazurin structural gene (pazS) is under the control of the Fnr/Nnr regulatory system, but no promoter-binding sequence could be recognized. The amino acid sequence of pseudoazurin from Paracoccus denitrificans is also reported.

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