Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29.
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.
D001412:Bacillus subtilis; D001435:Bacteriophages / Q000032:analysis*; D003001:Cloning, Molecular; D004261:DNA Replication; D011863:Radioimmunoassay; D014764:Viral Proteins / Q000276:immunology / Q000302:isolation & purification*; D014779:Virus Replication
D014764:Viral Proteins