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Early Intermediate State of Transforming Deoxyribonucleic Acid During Uptake by Bacillus subtilis¹

^a Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520

ABSTRACT

Examination of the effects of ethylenediaminetetraacetic acid (EDTA) on uptake of transforming deoxyribonucleic acid (DNA) by Bacillus subtilis allowed definition of a new intermediate state of transforming DNA during uptake. Markers in this state, termed "EDTA-resistant," are bound to the cell, are sensitive to inactivation by deoxyribonuclease, but may become deoxyribonuclease-resistant despite the addition of excess EDTA. Markers become EDTA-resistant quadratically with time after the addition of DNA to a competent culture, but linearly after exposure of competent cells to a brief pulse of DNA. An attractive model consistent with these findings is that EDTA blocks only the initiation of entry of a molecule of DNA.

² Present address: Department of Biochemistry, Duke University Medical Center, Durham, N.C. 27706.

¹ Part of a thesis submitted to Yale University in partial fulfillment of the requirements for the Ph.D. degree.