Summary
From ϕ29-infected Bacillus subtilis cells, we have isolated a protein fraction which promotes in vitro replication of ϕ29 DNA. This fraction catalyses both initiation and elongation, indicating that it contains the product of gene 3 (tp: terminal protein) and the product of gene 2 (gp2: probably a DNA polymerase), since initiation requires the two products (Blanco et al. 1983; Matsumoto et al. 1983). The fractions isolated from cells infected with temperature-sensitive (ts) mutants of gene 2 and gene 3 were thermolabile in both the initiation and elongation assays. When the pre-initiated material from the ts fractions of each mutant was heat-inactivated and mixed no complementation, restoring the elongation activity, was found. These results indicate: (i) tp and gp2 participate not only in the initiation but also in the elongation of ϕ29 DNA replication, (ii) they probably function in tight physical association with each other.
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Abbreviations
- gp2:
-
product of gene 2
- tp:
-
terminal protein
- DNAtp:
-
DNA with terminal protein covalently linked at both the 5′ ends
- ddCTP:
-
2′,3′-dideoxycytidine 5′-triphosphate
- ddGTP:
-
2′,3′-dideoxyguanosine 5′ triphosphate
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Communicated by M. Takanami
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Matsumoto, K., Saito, T., Kim, C.I. et al. Bacteriophage ϕ29 DNA replication in vitro: Participation of the terminal protein and the gene 2 product in elongation. Molec. Gen. Genet. 196, 381–386 (1984). https://doi.org/10.1007/BF00436183
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DOI: https://doi.org/10.1007/BF00436183