Transposon mutagenesis in Desulfovibrio desulfuricans: development of a random mutagenesis tool from Tn7.
The transposons Tn5, Tn7, Tn9, and Tn10 or their derivatives have been examined for transposition in the sulfate-reducing bacterium Desulfovibrio desulfuricans G20. Tn7 inserted with a frequency of 10(-4) to 10(-3) into a unique attachment site that shows strong homology with those sites identified in other gram-negative bacteria. Inactivation of the tnsD gene in Tn7, encoding the function directing insertion into the unique site, yielded a derivative that transposed essentially randomly with a frequency of ca. 10(-6) per donor. Derivatives of Tn5, but not wild-type Tn5, were also found to undergo random transposition at a similar frequency. No evidence was obtained for transposition of Tn9 or Tn10.
Wall
J D
JD
Biochemistry Department, University of Missouri-Columbia 65211, USA. Judy_D._Wall@muccmail.missouri.edu
Murnan
T
T
Argyle
J
J
English
R S
RS
Rapp-Giles
B J
BJ
D001483:Base Sequence; D003227:Conjugation, Genetic / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D003901:Desulfovibrio / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid
D004251:DNA Transposable Elements; D004269:DNA, Bacterial