Transposon Mutagenesis in Streptococcus Species
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Transposon Mutagenesis in Streptococcus Species. / Nilsson, Martin; Givskov, Michael; Tolker-Nielsen, Tim.
Microbial Transposon Mutagenesis: Protocols and Applications. Humana Press, 2019. s. 39-49 (Methods in Molecular Biology, Bind 2016).Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
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TY - CHAP
T1 - Transposon Mutagenesis in Streptococcus Species
AU - Nilsson, Martin
AU - Givskov, Michael
AU - Tolker-Nielsen, Tim
PY - 2019
Y1 - 2019
N2 - Mutant libraries, generated by transposons and screened for various phenotypes, have led to many important discoveries regarding gene functions in various organisms. In this chapter we describe the use of plasmid pMN100, a transposon vector constructed to perform in vivo transposition primarily in oral streptococci. Compared to in vitro transposition systems the conditional replicative features of the plasmid, and the inducible expression of the mariner Himar1 transposase, makes pMN100 particularly useful for bacterial strains showing a low transformation frequency. We outline how to transform plasmid pMN100 into Streptococcus mutans, carry out transposon mutagenesis, and determine the chromosomal location of inserted transposons. It is our prospect that the protocols can be used as guidelines for transposon mutagenesis in S. mutans as well as other species of streptococci.
AB - Mutant libraries, generated by transposons and screened for various phenotypes, have led to many important discoveries regarding gene functions in various organisms. In this chapter we describe the use of plasmid pMN100, a transposon vector constructed to perform in vivo transposition primarily in oral streptococci. Compared to in vitro transposition systems the conditional replicative features of the plasmid, and the inducible expression of the mariner Himar1 transposase, makes pMN100 particularly useful for bacterial strains showing a low transformation frequency. We outline how to transform plasmid pMN100 into Streptococcus mutans, carry out transposon mutagenesis, and determine the chromosomal location of inserted transposons. It is our prospect that the protocols can be used as guidelines for transposon mutagenesis in S. mutans as well as other species of streptococci.
KW - In vivo transposon mutagenesis
KW - Mariner
KW - pMN100
KW - Streptococci
U2 - 10.1007/978-1-4939-9570-7_4
DO - 10.1007/978-1-4939-9570-7_4
M3 - Book chapter
C2 - 31197707
AN - SCOPUS:85067452747
SN - 978-1-4939-9569-1
T3 - Methods in Molecular Biology
SP - 39
EP - 49
BT - Microbial Transposon Mutagenesis
PB - Humana Press
ER -
ID: 226876413