Insertional mutagenesis of an industrial strain of Streptococcus thermophilus.
Random mutagenesis of an industrial strain of Streptococcus thermophilus was achieved through an adapted version of a two-plasmid system. The mutagenesis strategy is based on random integration of derivatives of the non-replicative (Rep(-)) plasmid pORI19 by means of homologous recombination following a temperature shift that eliminates replication of the temperature-sensitive (Rep(ts)) helper plasmid pVE6007. In this way mutants were generated which were affected in bacteriophage sensitivity or sucrose metabolism. Homologues were identified of a protein related to folate metabolism from a bacteriophage-resistant mutant and of two subunits of an oligopeptide transport system from a mutant deficient in sucrose utilisation.
Labarre
C
C
National Food Biotechnology Centre, University College Cork, Ireland.
Schirawski
J
J
van der Zwet
A
A
Fitzgerald
G F
GF
van Sinderen
D
D
D000818:Animals; D001692:Biological Transport; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology; D008892:Milk / Q000382:microbiology; D016254:Mutagenesis, Insertional; D009842:Oligopeptides / Q000378:metabolism; D010957:Plasmids; D011995:Recombination, Genetic; D013291:Streptococcus / Q000235:genetics*; D017108:Streptococcus Phages; D013395:Sucrose / Q000378:metabolism
D009842:Oligopeptides; D013395:Sucrose