Translesion synthesis (TLS) is an event to cope with DNA damages. During TLS, the responsible TLS polymerase frequently elicits untargeted mutagenesis as potentially a source of genetic diversity. Identifying such untargeted mutations is challenging due to the bulk of DNA that does not undergo TLS. Here, we present a protocol to enrich a plasmid pool that underwent Pol V-mediated TLS in for mass sequencing. The concept of this protocol could be applied into any species. For complete details on the use and execution of this protocol, please refer to Isogawa et al. (2018).
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http://dx.doi.org/10.1016/j.xpro.2021.100399 | DOI Listing |
J Agric Food Chem
December 2024
The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
Ectoine is a valuable compatible solute with extensive applications in bioengineering, cosmetics, medicine, and the food industry. While certain halophilic bacteria can naturally produce ectoine, as a model organism for biomanufacturing, offers significant advantages to be engineered for potentially high-level ectoine production. However, complex metabolic flux distributions and byproduct formation present bottlenecks that limit ectoine production in .
View Article and Find Full Text PDFMicrobiol Res
December 2024
Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, United States. Electronic address:
Ehrlichia chaffeensis is an obligatory intracellular bacterium that infects monocytes and macrophages and causes human monocytic ehrlichiosis. Ehrlichia translocated factor-3 (Etf-3) is a type IV secretion system effector that binds host-cell ferritin light chain and induces ferritinophagy, thus increasing cellular labile iron pool for Ehrlichia proliferation. To further characterize roles of Etf-3 in Ehrlichia infection, we produced immune libraries of Etf-3-specific nanobodies (Nbs).
View Article and Find Full Text PDFJ Vis Exp
November 2024
Department of Chemistry, Institute of Biochemistry, BOKU University; CD Laboratory for Next Generation CAR T Cells;
Protein engineering enables the improvement of existing functions of a given protein or the generation of novel functions. One of the most widely used and versatile tools in the protein engineering field is yeast surface display, where a pool of randomized proteins is expressed on the surface of yeast. The linkage of phenotype (e.
View Article and Find Full Text PDFJ Appl Physiol (1985)
October 2024
Muscle Health Research Centre, School of Kinesiology and Health Science, York University, Toronto, Ontario, M3J 1P3, Canada.
Sci Rep
October 2024
Bioprocess Laboratory, Department of Biosystems Science and Engineering, ETH Zurich, 4056, Basel, Switzerland.
Transfer ribonucleic acids (tRNAs) are essential for protein synthesis, decoding mRNA sequences into amino acids. In E. coli K-12 MG1655, 86 tRNA genes are organized in 43 transcription units (TUs) and the essentiality of individual tRNA TUs in bacterial physiology remains unclear.
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