@inbook{140070aeb7ae4ae79d9a83f935b9f140,
title = "En passant mutagenesis: A Two Step Markerless Red Recombination System",
abstract = "Bacterial artificial chromosomes are used to maintain and modify large sequences of different origins in Escherichia coli. In addition to RecA-based shuttle mutagenesis, Red recombination is commonly used for sequence modification. Since foreign sequences, such as antibiotic resistance genes as well as frt- or loxP-sites are often unwanted in mutant BAC clones, we developed a Red-based technique that allows for the scarless generation of point mutations, deletions, and insertion of smaller and larger sequences. The method employs a sequence duplication that is inserted into the target sequence in the first recombination step and the excision of the selection marker by in vivo I-SceI cleavage and the second Red recombination. To allow for convenient and highly efficient mutagenesis without the use of additional plasmids, the E. coli strain GS1783 with a chromosomal encoded inducible Red- and I-SceI-expression was created. {\textcopyright} 2010 Springer Science+Business Media, LLC.",
keywords = "BAC, En passant mutagenesis, I-SceI, Markerless, Red recombination, BAC, En passant mutagenesis, I-SceI, Markerless, Red recombination, BAC, En passant mutagenesis, I-SceI, Markerless, Red recombination",
author = "{Karsten Tischer}, B. and Smith, {Gregory A.} and Nikolaus Osterrieder",
year = "2010",
doi = "10.1007/978-1-60761-652-8_30",
language = "English",
isbn = "9781607616511",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "421--430",
editor = "Jeff Braman",
booktitle = "In Vitro Mutagenesis Protocols",
address = "United States",
edition = "3rd",
}