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Measuring Homologous Recombination Rates between Chromosomal Locations in Salmonella
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
2019 (English)In: BIO-PROTOCOL, ISSN 2331-8325, Vol. 9, no 3, article id e3159Article in journal (Refereed) Published
Abstract [en]

Homologous recombination between two similar DNA molecules, plays an important role in the repair of double-stranded DNA breaks. Recombination can occur between two sister chromosomes, or between two locations of similar sequence identity within the same chromosome. The assay described here is designed to measure the rate of homologous recombination between two locations with sequence similarity within the same bacterial chromosome. For this purpose, a selectable/counter-selectable genetic cassette is inserted into one of the locations and homologous recombination repair rates are measured as a function of recombinational removal of the inserted cassette. This recombinational repair process is called gene conversion, non-reciprocal recombination. We used this method to measure the recombination rates between genes within gene families and to study the stability of mobile genetic elements inserted into members of gene families.

Place, publisher, year, edition, pages
BIO-PROTOCOL , 2019. Vol. 9, no 3, article id e3159
Keywords [en]
Homologous recombination, Homologous recombination rate(s), DNA repair, Double stranded DNA breaks, Salmonella, Gene families
National Category
Genetics
Identifiers
URN: urn:nbn:se:uu:diva-377594DOI: 10.21769/BioProtoc.3159ISI: 000458033700009OAI: oai:DiVA.org:uu-377594DiVA, id: diva2:1292940
Funder
Swedish Research Council, 2016-04449Swedish Research Council, 2017-03953Carl Tryggers foundation , CTS16: 194Carl Tryggers foundation , CTS17: 204Available from: 2019-03-01 Created: 2019-03-01 Last updated: 2019-03-07Bibliographically approved

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Brandis, GerritCao, ShaHughes, Diarmaid

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