New Insight into Plasmid-Driven T7 RNA Polymerase in Escherichia coli and Use as a Genetic Amplifier for a Biosensor.

ACS Synthetic Biology
Shih-I Tan, I-Son Ng

Abstract

T7 RNA polymerase (T7RNAP) and T7 promoter are powerful genetic components, thus a plasmid-driven T7 (PDT7) genetic circuit could be broadly applied for synthetic biology. However, the limited knowledge of the toxicity and instability of such a system still restricts its application. Herein, we constructed 16 constitutive genetic circuts of PDT7 and investigated the orthogonal effects in toxicity and instability. The T7 toxicity was elucidated from the construction processes and cell growth characterization, showing the importance of optimal orthogonality for PDT7. Besides, a protein analysis was performed to validate how the T7 system affected cell metabolism and led to the instability. The application of constitutive PDT7 in functional protein expressions, including carbonic anhydrase, lysine decarboxylase, and 5-ALA synthetase was demonstrated. Furthermore, PDT7 working as a genetic amplifier had been designed for E. coli cell-based biosensors, which illustrated the opportunities in the future of PDT7 used in synthetic biology.

References

May 5, 1986·Journal of Molecular Biology·F W Studier, B A Moffatt
Apr 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·P DavanlooF W Studier
Dec 22, 1999·Science·G M Cheetham, T A Steitz
Nov 30, 2000·Proceedings of the National Academy of Sciences of the United States of America·D TemiakovW T McAllister
Jul 28, 2009·Nature Biotechnology·Keith E J TyoGregory Stephanopoulos
Jun 30, 2012·Nucleic Acids Research·Karsten TemmeChristopher A Voigt
Feb 12, 2013·Nature Biotechnology·Piro SiutiTimothy K Lu
Mar 13, 2013·Proceedings of the National Academy of Sciences of the United States of America·David L Shis, Matthew R Bennett
Nov 26, 2013·Current Opinion in Chemical Biology·Alec A K NielsenChristopher A Voigt
Aug 1, 2014·Molecular Systems Biology·Thomas H Segall-ShapiroChristopher A Voigt
Jul 18, 2015·Nature Communications·Manish Kushwaha, Howard M Salis
Nov 26, 2015·Proceedings of the National Academy of Sciences of the United States of America·Shimyn SlomovicJames J Collins
Nov 28, 2016·Metabolic Engineering·Han ZhaoGuo-Qiang Chen
Feb 14, 2017·Nature Chemical Biology·Jinyue PuBryan C Dickinson
Oct 19, 2017·World Journal of Microbiology & Biotechnology·Zhen KangJian Chen
Feb 8, 2018·Current Opinion in Biotechnology·Hyun Gyu LimGyoo Yeol Jung
Mar 20, 2018·Nature Biotechnology·Thomas H Segall-ShapiroChristopher A Voigt
Jun 5, 2018·Metabolic Engineering·Chae Won KangGyoo Yeol Jung
Oct 6, 2018·Biotechnology Advances·Wenya WangRobert J Linhardt

❮ Previous
Next ❯

Citations

Jun 29, 2021·Biotechnology and Bioengineering·Yuanchao QianTing Lu

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

© 2021 Meta ULC. All rights reserved