Infection of a strain lysogenic for bacteriophage P1 CM with P1::Tn5 followed by simultaneous selection for the chloroamphenicol resistance associated with the resident prophage and the kanamycin resistance associated with Tn5 results in a large number of independent Tn5 insertion mutations. This superinfection-selection protocol is a fast, easy, and safe way to isolate null mutations in enteric bacteria without generating unwanted cryptic mutations elsewhere in the genome.
Download full-text PDF |
Source |
---|---|
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC239691 | PMC |
http://dx.doi.org/10.1128/aem.47.2.436-438.1984 | DOI Listing |
Metab Eng
January 2025
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kongens Lyngby, Denmark. Electronic address:
Advanced genome engineering enables precise and customizable modifications of bacterial species, and toolsets that exhibit broad-host compatibility are particularly valued owing to their portability. Tn5 transposon vectors have been widely used to establish random integrations of desired DNA sequences into bacterial genomes. However, the iteration of the procedure remains challenging because of the limited availability and reusability of selection markers.
View Article and Find Full Text PDFSci Rep
November 2024
Department of Food Hygiene and Consumer Health Protection, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
Campylobacter jejuni is a major cause of food- and water-borne bacterial infections in humans. A key factor helping bacteria to survive adverse environmental conditions is biofilm formation ability. Nonetheless, the molecular basis underlying biofilm formation by C.
View Article and Find Full Text PDFACS Synth Biol
November 2024
State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong 266237, People's Republic of China.
The efficiency of valuable metabolite production by engineered microorganisms underscores the importance of stable and controllable gene expression. While plasmid-based methods offer flexibility, integrating genes into host chromosomes can establish stability without selection pressure. However, achieving site-directed multicopy integration presents challenges, including site selection and stability.
View Article and Find Full Text PDFbioRxiv
September 2024
Center for Spatial and Functional Genomics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Res Sq
September 2024
Department of Food Hygiene and Consumer Health Protection, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.
is a major cause of food- and water-borne bacterial infections in humans. A key factor helping bacteria to survive adverse environmental conditions is biofilm formation ability. Nonetheless, the molecular basis underlying biofilm formation by remains poorly understood.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!