Transposase-Mediated Chromosomal Integration of Exogenous Genes in Acidithiobacillus ferrooxidans

Applied and Environmental Microbiology
Yuta InabaScott Banta

Abstract

The development of Acidithiobacillus ferrooxidans as a non-model host organism for synthetic biology is hampered by a lack of genetic tools and techniques. New plating and liquid-based selection methods were developed to improve the identification of transformed cell lines. Enabled by these methods, a hyperactive transposase was used to generate mutants with integrated genes for the expression of the superfolder green fluorescent protein (sfGFP) gene or a 2-keto decarboxylase (KDC) gene, which enabled the production and secretion of isobutyric acid (IBA). An inverse PCR method was used to identify the insertion sites of the KDC gene in several mutants, leading to the identification of a region on the chromosome that may be suitable for future genetic insertions. These results demonstrate that functional exogenous metabolic genes have been chromosomally integrated into A. ferrooxidans, and this advance will facilitate the future development of these cells for new biotechnology applications.IMPORTANCEAcidithiobacillus ferrooxidans is an iron- and sulfur-oxidizing chemolithoautotroph and is a key member of the microbial consortia used in industrial biomining applications. There is interest in exploiting these cells for other metal...Continue Reading

References

May 1, 1994·Journal of Bacteriology·J B PengX Z Bao
Mar 1, 1993·Nature Genetics·W Gish, D J States
Apr 29, 1998·The Journal of Biological Chemistry·I Y Goryshin, W S Reznikoff
Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·I Y GoryshinW S Reznikoff
Sep 6, 2000·Letters in Applied Microbiology·G HuangR G Birch
Jul 27, 2002·Annual Review of Microbiology·Douglas E Rawlings
Sep 24, 2002·Nature Biotechnology·Byung Jo YuSun Chang Kim
Jul 21, 2004·Journal of Bacteriology·Yisheng KangFrederick R Blattner
Aug 6, 2008·Annual Review of Genetics·William S Reznikoff
Feb 24, 2011·BMC Microbiology·Esteban Martínez-GarcíaVíctor de Lorenzo
Nov 1, 2011·Plasmid·Wesley Loftie-Eaton, Douglas E Rawlings
Nov 5, 2011·Environmental Microbiology·Violaine Bonnefoy, David S Holmes
Feb 9, 2012·Mobile DNA·Brian GreenBrendan P Cormack
Nov 13, 2014·Frontiers in Bioengineering and Biotechnology·Esteban Martínez-GarcíaPablo I Nikel
Jul 16, 2015·Biotechnology and Bioengineering·Timothy KernanScott Banta
Nov 23, 2016·Biotechnology and Applied Biochemistry·Timothy KernanScott Banta
Apr 4, 2017·Current Opinion in Biotechnology·Indrani BanerjeeScott Banta

❮ Previous
Next ❯

Citations

Jan 25, 2020·Nature Chemical Biology·Sarah ThorwallIan Wheeldon
Jan 27, 2021·Biotechnology for Biofuels·Lauren A Riley, Adam M Guss

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.

© 2022 Meta ULC. All rights reserved