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. 1974 Sep;71(9):3545–3549. doi: 10.1073/pnas.71.9.3545

Packaging and Maturation of DNA of Bacteriophage T7 In Vitro

Carol Kerr 1,2, Paul D Sadowski 1,2
PMCID: PMC433811  PMID: 4530321

Abstract

We have developed an in vitro complementation assay to demonstrate packaging and maturation of DNA of phage T7. Cells of Escherichia coli B infected with an appropriate T7 amber mutant are concentrated 200-fold and lysed by freezing and thawing. Two extracts from cells infected with different amber mutants are mixed and incubated at 30°. Positive complementation results in a 100-fold increase in phage titer.

Using this assay we have demonstrated the packaging of phage DNA from an extract that contains no phage heads (gene 9-, 10-), within head structures present in an extract that contains no phage DNA (gene 5-). We have also demonstrated an activity in extracts that contain no phage DNA or heads (gene 5-, 9-, 10-), which complements gene 19--infected cells. We have proven that this activity is due to the gene-19 product by showing that the activity is temperature-sensitive if the extract is made from cells infected with a mutant having a temperature-sensitive mutation in gene 19. This assay should be useful in elucidating the mechanism of packaging and maturation of DNA of phage T7.

Keywords: morphogenesis, in vitro complementation

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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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