Overproduction and purification of protein P6 of Bacillus subtilis phage phi 29: role in the initiation of DNA replication.
A phi 29 DNA fragment containing gene 6, required for DNA replication, has been cloned in plasmid pPLc28 under the control of the PL promoter of phage lambda. A polypeptide with an electrophoretic mobility close to that of p6 was labelled with 35S-methionine after heat induction. This protein, representing about 4% of the total E. coli protein after 1 h of induction, was obtained in a highly purified form. The protein was characterized as p6 by amino acid analysis and NH2-and COOH-terminal sequence determination. Protein p6 has an apparent molecular weight of 23,600, suggesting that the native form of the protein is a dimer. The purified protein p6 stimulated the protein-primed initiation of phi 29 DNA replication when added to purified proteins p2 (phi 29-coded DNA polymerase) and p3 (terminal protein).
Pastrana
R
R
Lázaro
J M
JM
Blanco
L
L
García
J A
JA
Méndez
E
E
Salas
M
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001412:Bacillus subtilis; D010582:Bacteriophage lambda / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics*; D004261:DNA Replication; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009876:Operon; D010957:Plasmids; D014461:Ultracentrifugation; D014764:Viral Proteins / Q000302:isolation & purification*
D000596:Amino Acids; D014764:Viral Proteins