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• W1972731911 abstract "The DNA plus cell interaction leading to bacterial transformation is an orderly two-particle reaction, dependent upon the first power of cell and DNA concentration,1-4 and of the duration of the extracellular contact.3-' When two physically unlinked markers are incorporated into the same cell, the double transformations become proportional to the second power of DNA concentration4' or duration of exposure.5 The competitive power of other DNA6 and the kinetics3' I suggest an early reversible step, but DNA is eventually physically incorporated.8-'0 The DNA which enters pneumococcal cells is rapidly separated into two equal fractions, high-molecular and degraded,1' and the regions associated with the introduced marker activity appear to be large tracts carrying only a single strand from the introduced donor DNA.12 In Hemophilus'3 and Bacillus subtilis14 cells also, it seems that only one transforming DNA strand survives intact. The reproducibility of the maximal pneumococcal transforming activity for several markers, as well as of various linkages, in DNA prepared without deliberate shear, has suggested that the recovered material may have relatively regular points of rupture when separated from the presumably much longer bacterial chromosome. This could happen if there were fixed discontinuities or regions of relative lability in the chromosome. This report will show that there is a tendency for reproducible sequences of marker entry in transformation by pneumococcal DNA, suggesting that the molecules as isolated from the cells have to some degree uniform terminations. The study makes use of separated single strands of DNA, reactivated by complexing with unmarked homologous DNA. Methods and Materials.-Deoxycholate lysates of pneumococci were used for all preparations of DNA. The preparation of chloroform gel DNA (C) and the methods of transformation have been frequently described.3' 15 The marker strains were multiple mutants and transformants of wild type (R6, derived from R36A), and the competent strain was a single colony derivative (R1-26) of the same strain. Zone centrifugation was done with 1 uAg DNA in 5-20 per cent sucrose gradients in saline at 34,000 rpm for 3 hr in the cold in celluloid tubes, and fractions of 2 drops collected from the bottom in sterile transformation medium and biologically tested. DNA preparations of type C were denatured with 0.1 M NaOH at 250C, then neutralized at 0?C in concentrations of about 25 ug/ml and put on 1-cm columns containing 5 ml of MAK having 0.7 the quantity of methylated albumin previously specified.6 About 0.2-0.6 mg of denatured DNA in 0.6 or 0.7 M NaCl was used; A fractions came at about 0.85 M and B at about 1.0 M salt. Renaturation was in 1.5 M NaCl at 65?C for 30 min, followed by 1-2 hr cooling; biological activity was measured at concentrations giving linear response. Rescue of denatured DNA was accomplished by renaturation with a double quantity of wild-type DNA and measured per unit of marked DNA present, expressed as per cent of activity given by native marked DNA." @default.
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• W1972731911 date "1966-11-01" @default.
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• W1972731911 title "MANIFESTATION OF LINEAR ORGANIZATION IN MOLECULES OF PNEUMOCOCCAL TRANSFORMING DNA" @default.
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• W1972731911 doi "https://doi.org/10.1073/pnas.56.5.1441" @default.
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