Home LiteratureArticle Details
PMID: 3158884 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Overproduction and purification of protein P6 of Bacillus subtilis phage phi 29: role in the initiation of DNA replication.

Nucleic acids research ·Vol. 13 ·No. 9 ·1985-05-10 ·Pages 3083-100

Pastrana R, Lázaro JM, Blanco L, García JA, Méndez E, Salas M

Abstract

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).

MeSH Terms
Amino Acid Sequence Amino Acids/analysis Bacillus subtilis Bacteriophage lambda/genetics Bacteriophages/genetics DNA Replication Electrophoresis, Polyacrylamide Gel Molecular Weight Operon Plasmids Ultracentrifugation Viral Proteins/isolation & purification
Chemicals
Amino Acids Viral Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Pastrana R
Lázaro J M
Blanco L
García J A
Méndez E
Salas M
References (43)
43 references, click to expand
  1. Molecular weights of coliphages and coliphage DNA. I. Measurement of the molecular weight of bacteriophage T7 by high-speed equilibrium centrifugation.
    J Mol Biol. 1970 Dec 28;54(3):537-46 PMID: 4923668
  2. In vitro initiation of bacteriophage phi 29 and M2 DNA replication: genes required for formation of a complex between the terminal protein and 5'dAMP.
    Mol Gen Genet. 1983;191(1):26-30 PMID: 6310350
  3. Temperature-sensitive mutants affected in DNA synthesis in phage phi29 of Bacillus subtilis.
    Eur J Biochem. 1972 Dec 4;31(2):367-71 PMID: 4630506
  4. Minicells of Bacillus subtilis.
    J Bacteriol. 1973 May;114(2):860-73 PMID: 4196259
  5. Suppressor-sensitive mutants and genetic map of Bacillus subtilis bacteriophage phi 29.
    Virology. 1974 Nov;62(1):1-16 PMID: 4214301
  6. Bacillus subtilis phage phi29. Characterization of gene products and functions.
    Eur J Biochem. 1976 JUL 1;66(2):229-41 PMID: 820555
  7. Genetic analysis of bacteriophage phi 29 of Bacillus subtilis: integration and mapping of reference mutants of two collections.
    J Virol. 1976 Aug;19(2):495-500 PMID: 822174
  8. Assembly of Bacillus subtilis phage phe29. 2. Mutants in the cistrons coding for the non-structural proteins.
    Eur J Biochem. 1977 Feb 15;73(1):57-72 PMID: 402270
  9. Carboxypeptidase Y in sequence determination of peptides.
    Methods Enzymol. 1977;47:84-93 PMID: 927202
  10. Identification of the protein firmly bound to the ends of bacteriophage phi 29 DNA.
    Virology. 1978 Feb;84(2):279-92 PMID: 203093
  11. Characterization of a protein covalently linked to the 5' termini of the DNA of Bacillus subtilis phage phi29.
    J Mol Biol. 1978 Feb 25;119(2):269-91 PMID: 416224
  12. Genome-linked protein associated with the 5' termini of bacteriophage phi29 DNA.
    J Virol. 1978 Sep;27(3):776-83 PMID: 100608
  13. Bacteriophage phi29 terminal protein: its association with the 5' termini of the phi29 genome.
    J Virol. 1978 Dec;28(3):895-904 PMID: 731797
  14. Construction of plasmid cloning vehicles that promote gene expression from the bacteriophage lambda pL promoter.
    Gene. 1979 Jan;5(1):59-76 PMID: 372049
  15. RNA polymerase binding sites and transcription map of the DNA of Bacillus subtilis phage phi29.
    J Mol Biol. 1979 Feb 5;127(4):411-36 PMID: 107317
  16. Structure of replicating DNA molecules of Bacillus subtilis bacteriophage phi 29.
    J Virol. 1980 Apr;34(1):187-99 PMID: 6768899
  17. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  18. Template requirements for initiation of phage phi 29 DNA replication in vitro.
    Proc Natl Acad Sci U S A. 1984 Jan;81(1):80-4 PMID: 6320176
  19. A novel DNA polymerase induced by Bacillus subtilis phage phi 29.
    Nucleic Acids Res. 1983 Dec 10;11(23):8333-42 PMID: 6424095
  20. Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29.
    Proc Natl Acad Sci U S A. 1984 Mar;81(6):1639-43 PMID: 6424120
  21. Characterization and purification of a phage phi 29-encoded DNA polymerase required for the initiation of replication.
    Proc Natl Acad Sci U S A. 1984 Sep;81(17):5325-9 PMID: 6433348
  22. Cloning and expression of gene 2, required for the protein-primed initiation of the Bacillus subtilis phage phi 29 DNA replication.
    Gene. 1984 Jul-Aug;29(1-2):33-40 PMID: 6092229
  23. Cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of Bacillus subtilis phage phi 29.
    Gene. 1984 Oct;30(1-3):87-98 PMID: 6096227
  24. The protein covalently linked to the 5' termini of the DNA of Bacillus subtilis phage phi 29 is involved in the initiation of DNA replication.
    Virology. 1980 Jul 15;104(1):84-96 PMID: 6771916
  25. DNA replication of bacteriophage phi 29: characterization of the intermediates and location of the termini of replication.
    Virology. 1980 Jul 30;104(2):323-38 PMID: 7395108
  26. Bacteriophage phi 29 infection of Bacillus subtilis minicells.
    Mol Gen Genet. 1980;180(3):539-45 PMID: 6780760
  27. Nucleotide sequence at the termini of the DNA of Bacillus subtilis phage phi 29.
    Proc Natl Acad Sci U S A. 1981 Mar;78(3):1446-50 PMID: 6262800
  28. Purified lambda regulatory protein cII positively activates promoters for lysogenic development.
    Nature. 1981 Jul 9;292(5819):128-32 PMID: 6264321
  29. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda.
    Gene. 1981 Oct;15(1):81-93 PMID: 6271633
  30. Cloning and analysis of strong promoters is made possible by the downstream placement of a RNA termination signal.
    Proc Natl Acad Sci U S A. 1981 Aug;78(8):4936-40 PMID: 6946440
  31. Nucleotide sequences of transcription and translation initiation regions in Bacillus phage phi 29 early genes.
    J Biol Chem. 1982 Jan 25;257(2):1053-62 PMID: 6274853
  32. Structure of protein-containing replicative intermediates of Bacillus subtilis phage phi 29 DNA.
    Virology. 1982 Jan 15;116(1):1-18 PMID: 6801848
  33. In vitro complex formation between bacteriophage phi 29 terminal protein and deoxynucleotide.
    Biochem Biophys Res Commun. 1982 Apr 14;105(3):1031-6 PMID: 6807309
  34. Nucleotide sequence of the major early region of bacteriophage phi 29.
    Gene. 1982 Mar;17(3):323-35 PMID: 6809534
  35. In vitro replication of bacteriophage phi 29 DNA.
    Proc Natl Acad Sci U S A. 1982 Sep;79(17):5245-8 PMID: 6813856
  36. Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5'-dAMP.
    Proc Natl Acad Sci U S A. 1982 Sep;79(18):5522-6 PMID: 6813861
  37. High level synthesis in Escherichia coli of the Bacillus subtilis phage phi 29 proteins p3 and p4 under the control of phage lambda PL promoter.
    Nucleic Acids Res. 1982 Oct 11;10(19):5773-84 PMID: 6292851
  38. Adenovirus DNA replication in vitro: identification of a host factor that stimulates synthesis of the preterminal protein-dCMP complex.
    Proc Natl Acad Sci U S A. 1982 Nov;79(21):6438-42 PMID: 6216480
  39. Factors involved in the initiation of phage phi 29 DNA replication in vitro: requirement of the gene 2 product for the formation of the protein p3-dAMP complex.
    Nucleic Acids Res. 1983 Mar 11;11(5):1309-23 PMID: 6402761
  40. Cloning and expression in Escherichia coli of the gene coding for the protein linked to the ends of Bacillus subtilis phage phi 29 DNA.
    Gene. 1983 Jan-Feb;21(1-2):65-76 PMID: 6301951
  41. Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication.
    Gene. 1983 Apr;22(1):103-13 PMID: 6305768
  42. Adenovirus DNA replication in vitro: synthesis of full-length DNA with purified proteins.
    Proc Natl Acad Sci U S A. 1983 Jul;80(14):4266-70 PMID: 6308611
  43. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.
    Biochemistry. 1971 Jun 22;10(13):2606-17 PMID: 4326772
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.9.3083
Published
1985-05-10
Pages
3083-100
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341222
Subset
IM
Grants
NIGMS NIH HHS · 5 R01 GM27242-05 · United States
External Links
PubMed source DOI: 10.1093/nar/13.9.3083 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2