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. 1974 Feb;71(2):431–435. doi: 10.1073/pnas.71.2.431

Formation In Vitro of Infective Joint Molecules of λ DNA by T4 Gene-32 Protein

Wilfried Wackernagel 1,2,*, Charles M Radding 1,2
PMCID: PMC388020  PMID: 4521814

Abstract

Half molecules of λ DNA that had been partially digested by λ exonuclease to expose homologous single strands were rejoined by the action of T4 gene-32 protein at 37° in the presence of Mg++. Measurements of infectivity in su- spheroplasts and sedimentation in sucrose demonstrated the formation of sus+ joint molecules from two preparations of sheared λ DNA that carried sus mutations at opposite ends of the genome. The biological activity of joint molecules made by annealing at 75° was diminished by the addition of the gene-32 protein in the absence of Mg++, and largely restored by the subsequent addition of Mg++. The specific infectivity of joint molecules made by gene-32 protein at 37° was similar to that of joint molecules made by annealing at 75°. The experimental system described provides a possible model for simulating early steps in genetic recombination.

Keywords: genetic recombination in vitro, joining reaction, synapsis

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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