Chapter title |
Markerless Genome Editing in Competent Streptococci
|
---|---|
Chapter number | 14 |
Book title |
Oral Biology
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6685-1_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6683-7, 978-1-4939-6685-1
|
Authors |
Roger Junges, Rabia Khan, Yanina Tovpeko, Heidi A. Åmdal, Fernanda C. Petersen, Donald A. Morrison |
Editors |
Gregory J. Seymour, Mary P. Cullinan, Nicholas C.K. Heng |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 9 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 2 | 22% |
Other | 1 | 11% |
Professor | 1 | 11% |
Student > Bachelor | 1 | 11% |
Researcher | 1 | 11% |
Other | 1 | 11% |
Unknown | 2 | 22% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 3 | 33% |
Agricultural and Biological Sciences | 2 | 22% |
Immunology and Microbiology | 1 | 11% |
Chemistry | 1 | 11% |
Unknown | 2 | 22% |