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Proc. Natl. Acad. Sci. USA Vol. 76, No. 6, pp. 2866-2870, June 1979
 

Summary: Proc. Natl. Acad. Sci. USA
Vol. 76, No. 6, pp. 2866-2870, June 1979
Genetics
Physical map of chromosomal nitrogen fixation (nif) genes of
Klebsiella pneumoniae
(transposable elements/restriction mapping/recombinant DNA)
GERARD E. RIEDEL*, FREDERICK M. AUSUBEL*, AND FRANK C. CANNONt
*Department of Biology, Harvard University, Cambridge, Massachusetts 02138; and tAgricultural Research Council Unit of Nitrogen Fixation, University of
Sussex, Falmer, Brighton BN1 9QJ, United Kingdom
Communicated by A. M. Pappenheimer, Jr., March 1, 1979
ABSTRACT We describe a method for the rapid determi-
nation of the physical location of mutations caused by insertion
of transposable elements. We used this method to construct a
detailed physical map of the nitrogen fixation (nif) gene cluster
of Klebsiela pneumoniae and to correlate it with the genetic
map. Total cellular DNA was isolated from individual strains,
each carrying an insertion in 1 of 15 different nif genes. The
DNA was digested with a restriction endonuclease, fractionated
by agarose gel electrophoresis, denatured, and blotted onto ni-
trocellulose filter paper. The DNA on the filters was hybridized

  

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

 

Collections: Biology and Medicine