scholarly journals Engineering a dynamic, controllable infectivity switch in bacteriophage T7

2021 ◽  
Author(s):  
Chutikarn Chitboonthavisuk ◽  
Phil Thaddeus Huss ◽  
Huai Luo Chun ◽  
Mikayla Fernholz ◽  
Srivatsan Raman
Keyword(s):  
Binding Sites ◽  
Phage Genome ◽  
Fold Increase ◽  
Bacteriophage T7 ◽  
Transcriptional Repressors ◽  
Proof Of Concept ◽  
Simple Circuit ◽  
Ribosome Binding ◽  
Ribosome Binding Sites ◽  
Phage Activity

Transcriptional repressors play an important role in regulating phage genomes. Here, we examined how synthetic regulation based on repressors can be to create a dynamic, controllable infectivity switch in bacteriophage T7. We engineered T7 by replacing a large region of the early phage genome with combinations of ligand-responsive promoters and ribosome binding sites (RBS) designed to control the phage RNA polymerase. Phages with the engineered switch showed virulence comparable to wildtype when not repressed, indicating the phage can be engineered without a loss of fitness. When repressed, the most effective switch used a TetR promoter and a weak RBS, resulting in a two-fold increase in latent period (time to lyse host) and change in phage titer. Further, phage activity could be tuned by varying inducer concentrations. Our study provides a proof of concept for a simple circuit for user control over phage infectivity.

Design of Ribosome Binding Sites in Streptomyces coelicolor

2017 ◽  
Vol 14 (4) ◽  
Author(s):  
Hong-Dou Luo ◽  
Yang Tao ◽  
Wen-Guang Wang ◽  
Tao Lin ◽  
Yue-Yue Wang ◽  
...  
Keyword(s):  
Binding Sites ◽  
Streptomyces Coelicolor ◽  
Ribosome Binding ◽  
Ribosome Binding Sites

scholarly journals Tuning a bi-enzymatic cascade reaction in Escherichia coli to facilitate NADPH regeneration for ε-caprolactone production

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Jinghui Xiong ◽  
Hefeng Chen ◽  
Ran Liu ◽  
Hao Yu ◽  
Min Zhuo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Regeneration System ◽  
Nadph Regeneration ◽  
Whole Cell ◽  
Cyclohexanone Monooxygenase ◽  
Ribosome Binding ◽  
Ribosome Binding Sites ◽  
Whole Cell Biocatalyst ◽  
Substrate Tolerance

Abstractε-Caprolactone is a monomer of poly(ε-caprolactone) which has been widely used in tissue engineering due to its biodegradability and biocompatibility. To meet the massive demand for this monomer, an efficient whole-cell biocatalytic approach was constructed to boost the ε-caprolactone production using cyclohexanol as substrate. Combining an alcohol dehydrogenase (ADH) with a cyclohexanone monooxygenase (CHMO) in Escherichia coli, a self-sufficient NADPH-cofactor regeneration system was obtained. Furthermore, some improved variants with the better substrate tolerance and higher catalytic ability to ε-caprolactone production were designed by regulating the ribosome binding sites. The best mutant strain exhibited an ε-caprolactone yield of 0.80 mol/mol using 60 mM cyclohexanol as substrate, while the starting strain only got a conversion of 0.38 mol/mol when 20 mM cyclohexanol was supplemented. The engineered whole-cell biocatalyst was used in four sequential batches to achieve a production of 126 mM ε-caprolactone with a high molar yield of 0.78 mol/mol.

Expression of synthetic calcitonin genes in plasmid vectors containing tandemly repeated non-overlapping ribosome binding sites

1989 ◽  
Vol 21 (9) ◽  
pp. 987-996 ◽  
Author(s):  
Krassimir Alexciev ◽  
Anna Uscheva ◽  
Maja Pavlova ◽  
Libert Yavachev ◽  
Ivan Ivanov
Keyword(s):  
Binding Sites ◽  
Plasmid Vectors ◽  
Ribosome Binding ◽  
Ribosome Binding Sites

scholarly journals Evidence for Context-Dependent Complementarity of Non-Shine-Dalgarno Ribosome Binding Sites to Escherichia coli rRNA

2013 ◽  
Vol 8 (5) ◽  
pp. 958-966 ◽  
Author(s):  
Pamela A. Barendt ◽  
Najaf A. Shah ◽  
Gregory A. Barendt ◽  
Parth A. Kothari ◽  
Casim A. Sarkar
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Ribosome Binding ◽  
Ribosome Binding Sites ◽  
Context Dependent

scholarly journals Multiple divergent picobirnaviruses with functional prokaryotic Shine-Dalgarno ribosome binding sites present in cloacal sample of a diarrheic chicken

Virology ◽  
2018 ◽  
Vol 525 ◽  
pp. 62-72 ◽  
Author(s):  
Ákos Boros ◽  
Beáta Polgár ◽  
Péter Pankovics ◽  
Hajnalka Fenyvesi ◽  
Péter Engelmann ◽  
...  
Keyword(s):  
Binding Sites ◽  
Ribosome Binding ◽  
Ribosome Binding Sites

Quantitativer Vergleich der Initiator-Regionen (Ribosome Binding Sites) von 12 aus Escherichia coli stammenden Nukleotidsequenzen (RNA-und DNA-Phagen), die aus Triplets zusammengesetzt sind / Quantitative Comparison of Ribosome Binding Sites of Twelve Nucleotide Sequences from Escherichia coli (RNA-and DNA Phages) Based on Triplet Patterns

1979 ◽  
Vol 34 (9-10) ◽  
pp. 797-804 ◽  
Author(s):  
Erich Köhler
Keyword(s):  
Escherichia Coli ◽  
Binding Sites ◽  
Binary Sequence ◽  
Simple Calculation ◽  
Phage Gene ◽  
Ribosome Binding ◽  
Ribosome Binding Sites ◽  
Pyrimidine Bases ◽  
Triplet Code ◽  
Rna Phage

Abstract The molecular structure of ribosome binding sites of ten phage genes and two messengers of Escherichia coli were compared concerning the signation parts which are presumably used by ribosomes for recognition and binding. With a simple calculation based on triplet patterns sofar unknown agreements between all of these sequences were found. In several cases it was shown that agreements between old sequences are easier reognizable if the purine-and pyrimidine bases are put into the triplets instead of the four A, G, C, and U (T) bases. In such cases “homologous” parts of sequences were recognized with more distinctness. This is true in our case for the double triplet (hexaplet) py-pu-pu-pu-pu-(pu) and the binding site triplet py-pu-pu, which are preceding the ini­tiator. These triplets are in specific positions in all twelve sequences which were compared. The different course of the quaternary and the binary conformity curves (diagram 1) may show for the investigated area that the RNA phage gene-part is organized according to the well known quater­nary triplet code. On the contrary the phage φ-gene-part seems to be organized according to a more simple, binary triplet sequence of purine and pyrimidine bases. The binary sequence seems to be the more original, the quaternary the derived one.

Sign in / Sign up

Export Citation Format

Share Document