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Construction and properties of recombinant plasmids containing the rII genes of bacteriophage T4

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Summary

The EcoRI digestion products of phage T4 DNA have been examined using a phage DNA transformation assay. A 2.6x106 Dalton fragment was found to contain the rII genes. This fragment was purified and then treated with HindIII endonuclease. The cleavage products were ligated to the vector plasmid pBR313 and viable recombinant plasmids recovered. A genetic assay was employed to demonstrate that the recombinants contained T4 DNA and to localize on the phage genetic map the EcoRI and HindIII sites cleaved during the construction of the plasmids. Preliminary characterization suggests that a fragment covering the beginning of the rIIA gene possibly contains a promotor which is active in uninfected cells.

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Abbreviations

Ap:

ampicillin

Tc:

tetracycline

Mdal:

106 Daltons

bp:

base pairs

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Communicated by W. Arber

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Selzer, G., Bolle, A., Krisch, H. et al. Construction and properties of recombinant plasmids containing the rII genes of bacteriophage T4. Molec. Gen. Genet. 159, 301–309 (1978). https://doi.org/10.1007/BF00268267

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