@inbook{d3c0ea457e814cb4900b099b5372e587,
title = "Genetic Methods and Construction of Chromosomal Mutations in Methanogenic Archaea",
abstract = "Genetic manipulation through markerless exchange enables the modification of several genomic regions without leaving a selection marker in the genome. Here, a method using hpt coding for hypoxanthine phosphoribosyltransferase as a counter selectable marker is described. For Methanosarcina species a chromosomal deletion of the hpt gene is firstly generated, which confers resistance to the purine analogue 8-aza-2,6-diaminopurine (8-ADP). In a second step, the reintroduction of the hpt gene on a plasmid leads to a selectable loss of 8-ADP resistance after a homologous recombination event (pop-in). A subsequent pop-out event restores the 8-ADP resistance and can generate chromosomal mutants with frequencies of about 50%.",
keywords = "Chromosomal mutant, Genetic manipulation, Genetic system, Markerless exchange, Methanogenic archaea, Pop-in and pop-out",
author = "Johanna Thomsen and Katrin Weidenbach and Metcalf, {William W.} and Schmitz, {Ruth A.}",
note = "Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-2445-6_6",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "105--117",
booktitle = "Methods in Molecular Biology",
}