Purification in a Functional Form of the Terminal Protein of Bacillus Subtilis Phage Phi 29

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1984 Mar 1

PMID 6424120

Citations 19

Authors

I Prieto

J M Lazaro

J A Garcia

J M Hermoso

M Salas

Affiliations

Soon will be listed here.

Abstract

Phage phi 29 terminal protein, p3, essentially pure, was isolated in a denatured form from viral particles, and anti-p3 antiserum was obtained. A radioimmunoassay to detect and quantitate protein p3 was developed. By using this assay, native protein p3 was highly purified from Escherichia coli cells harboring a gene 3-containing recombinant plasmid. After three purification steps, the protein was more than 96% pure; its amino acid composition was very similar to that deduced from the nucleotide sequence of gene 3. The purified protein was active in the formation of the covalent p3-dAMP initiation complex when supplemented with extracts of B. subtilis infected with a sus mutant of phi 29 in gene 3. No DNA polymerase or ATPase activities were present in the final preparation of protein p3.

Citing Articles

Structures of phi29 DNA polymerase complexed with substrate: the mechanism of translocation in B-family polymerases.

Berman A, Kamtekar S, Goodman J, Lazaro J, de Vega M, Blanco L EMBO J. 2007; 26(14):3494-505.

PMID: 17611604 PMC: 1933411. DOI: 10.1038/sj.emboj.7601780.

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.

Characterization and purification of a phage phi 29-encoded DNA polymerase required for the initiation of replication.

Blanco L, Salas M Proc Natl Acad Sci U S A. 1984; 81(17):5325-9.

PMID: 6433348 PMC: 391696. DOI: 10.1073/pnas.81.17.5325.

Replication of phage phi 29 DNA with purified terminal protein and DNA polymerase: synthesis of full-length phi 29 DNA.

Blanco L, Salas M Proc Natl Acad Sci U S A. 1985; 82(19):6404-8.

PMID: 3863101 PMC: 390724. DOI: 10.1073/pnas.82.19.6404.

Effect of NH4+ ions on phi 29 DNA-protein p3 replication: formation of a complex between the terminal protein and the DNA polymerase.

Blanco L, Prieto I, Gutierrez J, Bernad A, Lazaro J, Hermoso J J Virol. 1987; 61(12):3983-91.

PMID: 3682063 PMC: 256019. DOI: 10.1128/JVI.61.12.3983-3991.1987.

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