Molecular fate of DNA in genetic transformation of Pneumococcus.

Extracts of pneumococcal cells which had incorporated [ 32 P]DNA were fractionated in order to determine the fate of the incorporated radioactivity. Immediately after entry, about half of the DNA was found to be converted to a single-stranded form; the other half was degraded to dialysable oligonucleotides and inorganic phosphate. Very little native [ 32 P]DNA was present. On incubation of the cells for various lengths of time after introduction of the DNA, radioactivity was rapidly incorporated into the native DNA fraction. The sources of this radioactivity were (1) the fragments initially present which apparently were incorporated by means of normal synthetic processes, and (2) the single-stranded DNA some of which may have been transferred intact. This latter transfer appears to be the route by which genetic information is transferred. The increase of radioactivity in native DNA and the depletion of the single-stranded DNA corresponded in time with recovery of a genetic factor introduced by the donor DNA. The extent to which this genetic factor was recovered, determined both by direct measurement of its transforming activity and by measurement of the frequency of transformed cells, corresponded to a physical integration of about one-quarter of the donor DNA taken up by the cells.

[1]  A. D. Hershey,et al.  A fractionating column for analysis of nucleic acids. , 1960, Analytical biochemistry.

[2]  E. Chargaff,et al.  Nucleic Acids , 2020, Definitions.

[3]  G. Zubay A THEORY ON THE MECHANISM OF MESSENGER-RNA SYNTHESIS. , 1962, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Meselson,et al.  Chromosome brekage accompanying genetic recombination in bacteriophage. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Lacks,et al.  A study of the genetic material determining an enzyme activity in Pneumococcus , 1960 .

[6]  W. Wood,et al.  The effect of enzymatically synthesized ribonucleic acid on amino acid incorporation by a soluble protein-ribosome system from Escherichia coli. , 1962, Proceedings of the National Academy of Sciences of the United States of America.

[7]  W. R. Guild Transformation by denatured deoxyribonucleic acid. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Meselson,et al.  EQUILIBRIUM SEDIMENTATION OF MACROMOLECULES IN DENSITY GRADIENTS. , 1957, Proceedings of the National Academy of Sciences of the United States of America.

[9]  S. Goodgal,et al.  Recombination during transformation in Hemophilus influenzae. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[10]  L. Tolmach,et al.  Genetic transformation. I. Cellular incorporation of DNA accompanying transformation in Pneumococcus. , 1957, Biochimica et biophysica acta.

[11]  R. Hotchkiss CYCLICAL BEHAVIOR IN PNEUMOCOCCAL GROWTH AND TRANSFORMABILITY OCCASIONED BY ENVIRONMENTAL CHANGES. , 1954, Proceedings of the National Academy of Sciences of the United States of America.

[12]  S. Lacks,et al.  Formation of amylomaltase after genetic transformation of pneumococcus. , 1960, Biochimica et biophysica acta.

[13]  R. M. Herriott FORMATION OF HETEROZYGOTES BY ANNEALING A MIXTURE OF TRANSFORMING DNAS. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[14]  O. Avery,et al.  Studies on the chemical nature of the substance inducing transformation on pneumococcal types; an improved method for the isolation of the transforming substance and its application to Pneumococcus Types II, III, and VI. , 1946, The Journal of experimental medicine.

[15]  J. Weigle,et al.  Exchange of DNA in the recombination of bacteriophage lambda. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[16]  R. Hotchkiss,et al.  Fate of Transforming Deoxyribonucleate following Fixation by Transformable Bacteria: I , 1960, Nature.

[17]  P. Doty,et al.  STRAND SEPARATION AND SPECIFIC RECOMBINATION IN DEOXYRIBONUCLEIC ACIDS: PHYSICAL CHEMICAL STUDIES. , 1960, Proceedings of the National Academy of Sciences of the United States of America.

[18]  M. Fox,et al.  Deoxyribonucleic acid incorporation by transformed bacteria. , 1957, Biochimica et biophysica acta.

[19]  M. Roger,et al.  Selective heat inactivation of pneumococcal transforming deoxyribonucleate. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M Meselson,et al.  THE REPLICATION OF DNA IN ESCHERICHIA COLI. , 1958, Proceedings of the National Academy of Sciences of the United States of America.

[21]  P. Doty,et al.  The biological and physical heterogeneity of thermally denatured and renatured deoxyribonucleic acid. , 1961, Biochimica et biophysica acta.

[22]  L. Tolmach,et al.  Genetic transformation. II. The significance of damage to the DNA molecule. , 1959, Biochimica et biophysica acta.