Home

About

Advanced Search

Browse by Discipline

Scientific Societies

E-print Alerts

Add E-prints

E-print Network
FAQHELPSITE MAPCONTACT US


  Advanced Search  

 
INFECTION AND IMMUNITY, JUlY 1989, p. 2141-2148 Vol. 57, No. 7 0019-9567/89/072141-08$02.00/0
 

Summary: INFECTION AND IMMUNITY, JUlY 1989, p. 2141-2148 Vol. 57, No. 7
0019-9567/89/072141-08$02.00/0
Copyright 1989, American Society for Microbiology
General Method for Site-Directed Mutagenesis in Escherichia coli
018ac:K1:H7: Deletion of the Inducible Superoxide Dismutase
Gene, sodA, Does Not Diminish Bacteremia in Neonatal Rats
CRAIG A. BLOCH,lt* GRACE M. THORNE,2 AND FREDERICK M. AUSUBEL'
Department of Genetics, Harvard Medical School, and Department of Molecular Biology, Massachusetts General
Hospital, Boston, Massachusetts 02114,1 and Department ofPediatrics, Harvard Medical School, and Division
ofInfectious Diseases, Department ofMedicine, Children's Hospital, Boston, Massachusetts 021 j52
Received 4 January 1989/Accepted 10 April 1989
A defined deletion in the Escherichia coli K-12 sodA gene (encoding manganese-superoxide dismutase) linked
to a nontransposable selectable marker was generated by transposon TnS insertion in combination with in vitro
mutagenesis. This mutant allele was used to replace the wild-type sodA gene in an E. coli clinical isolate of
serotype 018ac:K1:H7 by bacteriophage P1 transduction. The 018ac:K1:H7 sodA mutant contained no
manganese-superoxide dismutase and no hybrid manganese-iron-superoxide dismutase. The sod4 mutant was
more sensitive to paraquat toxicity than were the parental strain and an isogenic mutant bearing an analogously
constructed sodA+ TnS insertion allele. In a suckling rat model for bacteremia following oral inoculation of E.
coli Kl, the sodA mutant was undiminished in its capabilities both to colonize the gastrointestinal tract and,
surprisingly, to cause bacteremia. In conjunction with the rat model for E. coli Kl pathogenesis, the method

  

Source: Ausubel, Frederick M. - Department of Genetics, Harvard University

 

Collections: Biology and Medicine