Construction and characterization of Streptococcus suis-Escherichia coli shuttle cloning vectors

Plasmid
Daisuke TakamatsuTsutomu Sekizaki

Abstract

pSSU1, a native plasmid of Streptococcus suis DAT1, was used to construct pSET-series shuttle vectors. In addition to the replication function of pSSU1, these vectors contain the multiple cloning sites and lacZ' gene from pUC19, which means that X-gal screening can be used to select recombinants in Escherichia coli. pSET1, pSET2, and pSET3 carry cat, spc, and both of these genes, respectively, as selectable markers. These vectors could be introduced into S. suis, E. coli, Salmonella typhimurium, S. pneumoniae, and S. equi ssp. equi by electrotransformation. The recA gene was cloned from S. suis and sequenced, and this information was used in the construction of a recA mutant of S. suis. Transformation frequencies and/or plasmid stability of all pSET vectors tested were decreased in both S. suis and E. coli recA mutants compared with the parental strains. These results suggested that functional RecA protein improved the maintenance of pSET vectors in both S. suis and E. coli. Moreover, cloning of the functional S. suis recA gene into pSET2 and complementation analysis of the recA mutant were successful in S. suis but not in E. coli. These results showed that pSET vectors are useful tools for cloning and analyzing S. suis genes i...Continue Reading

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Citations

Apr 16, 2013·Applied and Environmental Microbiology·Daisuke TakamatsuMakoto Osaki
Jun 4, 2013·Applied and Environmental Microbiology·Alexandra Faulds-Pain, Brendan W Wren
May 14, 2008·Infection and Immunity·Nahuel FittipaldiMarcelo Gottschalk
Mar 13, 2002·Journal of Bacteriology·Daisuke TakamatsuTsutomu Sekizaki
Jan 22, 2005·Journal of Bacteriology·Tsutomu SekizakiYoshihiro Shimoji
Sep 18, 2012·PloS One·Marie-Pier LecoursMarcelo Gottschalk
Nov 29, 2013·The Journal of Infectious Diseases·Dan TakeuchiKazunori Oishi
Nov 26, 2010·Microbial Pathogenesis·Tao WuHuanchun Chen
Jul 14, 2010·Microbial Pathogenesis·Peng LiShuzhang Feng
May 19, 2009·Veterinary Microbiology·Youhui SiHuanchun Chen
Nov 8, 2008·Molecular Microbiology·Nahuel FittipaldiMarcelo Gottschalk
Aug 16, 2011·Microbial Pathogenesis·Chen TanHuanchun Chen
Mar 7, 2003·FEMS Microbiology Letters·Makoto OsakiTsutomu Sekizaki

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