In Vitro Replication of Bacteriophage Phi 29 DNA

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Sep 1

PMID 6813856

Citations 15

Authors

K Watabe

M F Shih

A Sugino

J Ito

Affiliations

Soon will be listed here.

Abstract

We have been studying the mechanisms of linear DNA replication by using Bacillus bacteriophage phi 29 as a model system. To isolate and characterize the proteins required for phi 29 DNA replication, we have developed a cell-free replication system. A cell-free extract prepared from phi 29-infected Bacillus subtilis catalyzes the semiconservative replication of phi 29 DNA, but only if exogenous phi 29 DNA-protein complex is used as the template. This template consists of linear duplex DNA with a 30,000-dalton terminal protein attached covalently to both 5' ends. Replication starts nonsimultaneously at or near both ends of the template. The extract also catalyzes the specific binding between dATP and the phi 29 terminal protein. Thus, the in vitro system closely mimics the in vivo replication of phi 29 DNA. This system should allow characterization of the phi 29 DNA replication machinery.

Citing Articles

Bacteriophage phi 29 DNA replication in vitro: participation of the terminal protein and the gene 2 product in elongation.

Matsumoto K, Saito T, Kim C, Ando T, Hirokawa H Mol Gen Genet. 1984; 196(3):381-6.

PMID: 6438445 DOI: 10.1007/BF00436183.

Replication of bacteriophage phi 29 DNA in vitro: the roles of terminal protein and DNA polymerase.

Watabe K, Leusch M, Ito J Proc Natl Acad Sci U S A. 1984; 81(17):5374-8.

PMID: 6433349 PMC: 391706. DOI: 10.1073/pnas.81.17.5374.

Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29.

Prieto I, Lazaro J, Garcia J, Hermoso J, Salas M Proc Natl Acad Sci U S A. 1984; 81(6):1639-43.

PMID: 6424120 PMC: 344973. DOI: 10.1073/pnas.81.6.1639.

A novel DNA polymerase induced by Bacillus subtilis phage phi 29.

Watabe K, Ito J Nucleic Acids Res. 1983; 11(23):8333-42.

PMID: 6424095 PMC: 326585. DOI: 10.1093/nar/11.23.8333.

Protein-primed initiation of phage phi 29 DNA replication.

Watabe K, Shin M, Ito J Proc Natl Acad Sci U S A. 1983; 80(14):4248-52.

PMID: 6410387 PMC: 384014. DOI: 10.1073/pnas.80.14.4248.

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