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Methods Mol Biol

Title:		Markerless Genome Editing in Competent Streptococci
Author(s):		Junges R; Khan R; Tovpeko Y; Amdal HA; Petersen FC; Morrison DA;
Address:		"Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway. Department of Biological Sciences, College of Liberal Arts and Sciences, University of Illinois at Chicago, 845 West Taylor Street, (MC 066), Chicago, IL, 60607, USA. Department of Biological Sciences, College of Liberal Arts and Sciences, University of Illinois at Chicago, 845 West Taylor Street, (MC 066), Chicago, IL, 60607, USA. damorris@uic.edu"
Journal Title:		Methods Mol Biol
Year:		2017
Volume:		1537
Issue:
Page Number:		233 - 247
DOI:		10.1007/978-1-4939-6685-1_14
ISSN/ISBN:		1940-6029 (Electronic) 1064-3745 (Linking)
Abstract:		"Selective markers employed in classical mutagenesis methods using natural genetic transformation can affect gene expression, risk phenotypic effects, and accumulate as unwanted genes during successive mutagenesis cycles. In this chapter, we present a protocol for markerless genome editing in Streptococcus mutans and Streptococcus pneumoniae achieved with an efficient method for natural transformation. High yields of transformants are obtained by combining the unimodal state of competence developed after treatment of S. mutans with sigX-inducing peptide pheromone (XIP) in a chemically defined medium (CDM) or of S. pneumoniae with the competence-stimulating peptide (CSP) together with use of a donor amplicon carrying extensive flanking homology. This combination ensures efficient and precise integration of a new allele by the recombination machinery present in competent cells"
Keywords:		"Bacterial Proteins/metabolism/pharmacology *Gene Editing Genes, Bacterial *Genetic Markers *Genome, Bacterial Mutation Sequence Deletion Sigma Factor/metabolism/pharmacology Streptococcus/*genetics Transformation, Bacterial/drug effects Csp Competence Mar;"
Notes:		"MedlineJunges, Roger Khan, Rabia Tovpeko, Yanina Amdal, Heidi A Petersen, Fernanda C Morrison, Donald A eng Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. 2016/12/08 Methods Mol Biol. 2017; 1537:233-247. doi: 10.1007/978-1-4939-6685-1_14"