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- W2335593640 abstract "A method is described for assessing the biological activity of DNA isolated from the bacteriophage T4. Genetic transformation of mutant phages by wild-type DNA is the underlying principle of this system. The method involves infection of the host Escherichia coli B with a mutant phage along with DNA isolated from a wild-type phage, resulting in the appearance of wild-type phages in the progeny. To this end it was necessary to remove the cell wall of the host; this was achieved by converting the cells into spheroplasts by means of the penicillin method. The phage mutant to be transformed was converted, by treatment with urea, into a form that is able to infect spheroplasts (the so-called π-particles). The efficiency (the number of transformants per DNA molecule) was 6 · 10−5/B, in which B represents the mean burst size of the transformants. Some parameters of this system were studied: the optimum concentration of π-particles, the relation between DNA concentration and yield of transformants, the kinetics and burst sizes. Amber mutants can also be used in this system, provided their reversion frequency is low; the expression of some function of the wild-type DNA added is required for the production of phage progeny in the non-permissive host. Using transforming DNA with a marker affecting the plaque morphology (DNA isolated from an rI mutant) the simultaneous transfer of two markers was studied." @default.
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- W2335593640 date "1968-06-01" @default.
- W2335593640 modified "2023-09-27" @default.
- W2335593640 title "Genetic transformation of the bacteriophage T4" @default.
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- W2335593640 doi "https://doi.org/10.1016/0005-2787(68)90297-9" @default.
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