Development of an in vitro bacteriophage N4 DNA replication system

J. K. Rist, Margaret S Pearle, A. Sugino, L. B. Rothman-Denes

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

An in vitro DNA replication system for bacteriophage N4-infected Escherichia coli has been developed. It requires MgCl2, all four deoxyribonucleoside triphosphates, and exogenously added N4 phage DNA; other DNAs are used inefficiently or not at all. Ribonucleoside triphosphates are not required, although they stimulate DNA synthesis. In vitro replication starts at the ends of the N4 genome and moves progressively inward. Initiation occurs through hairpin priming at the 3' end of the genome, but shows a strong preference for the right end. Three N4 gene products (dnp, dbp, and exo) required in vivo for N4 DNA synthesis are absolutely required in the in vitro system. These findings are discussed with respect to the mode of N4 DNA replication.

Original languageEnglish (US)
Pages (from-to)10506-10510
Number of pages5
JournalJournal of Biological Chemistry
Volume261
Issue number23
StatePublished - 1986

Fingerprint

Bacteriophage N4
Bacteriophages
DNA Replication
DNA
Genes
Genome
Deoxyribonucleosides
Ribonucleosides
Magnesium Chloride
Escherichia coli
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rist, J. K., Pearle, M. S., Sugino, A., & Rothman-Denes, L. B. (1986). Development of an in vitro bacteriophage N4 DNA replication system. Journal of Biological Chemistry, 261(23), 10506-10510.

Development of an in vitro bacteriophage N4 DNA replication system. / Rist, J. K.; Pearle, Margaret S; Sugino, A.; Rothman-Denes, L. B.

In: Journal of Biological Chemistry, Vol. 261, No. 23, 1986, p. 10506-10510.

Research output: Contribution to journalArticle

Rist, JK, Pearle, MS, Sugino, A & Rothman-Denes, LB 1986, 'Development of an in vitro bacteriophage N4 DNA replication system', Journal of Biological Chemistry, vol. 261, no. 23, pp. 10506-10510.
Rist, J. K. ; Pearle, Margaret S ; Sugino, A. ; Rothman-Denes, L. B. / Development of an in vitro bacteriophage N4 DNA replication system. In: Journal of Biological Chemistry. 1986 ; Vol. 261, No. 23. pp. 10506-10510.
@article{92b25a65f2e040bf8cc114d027d8b272,
title = "Development of an in vitro bacteriophage N4 DNA replication system",
abstract = "An in vitro DNA replication system for bacteriophage N4-infected Escherichia coli has been developed. It requires MgCl2, all four deoxyribonucleoside triphosphates, and exogenously added N4 phage DNA; other DNAs are used inefficiently or not at all. Ribonucleoside triphosphates are not required, although they stimulate DNA synthesis. In vitro replication starts at the ends of the N4 genome and moves progressively inward. Initiation occurs through hairpin priming at the 3' end of the genome, but shows a strong preference for the right end. Three N4 gene products (dnp, dbp, and exo) required in vivo for N4 DNA synthesis are absolutely required in the in vitro system. These findings are discussed with respect to the mode of N4 DNA replication.",
author = "Rist, {J. K.} and Pearle, {Margaret S} and A. Sugino and Rothman-Denes, {L. B.}",
year = "1986",
language = "English (US)",
volume = "261",
pages = "10506--10510",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "23",

}

TY - JOUR

T1 - Development of an in vitro bacteriophage N4 DNA replication system

AU - Rist, J. K.

AU - Pearle, Margaret S

AU - Sugino, A.

AU - Rothman-Denes, L. B.

PY - 1986

Y1 - 1986

N2 - An in vitro DNA replication system for bacteriophage N4-infected Escherichia coli has been developed. It requires MgCl2, all four deoxyribonucleoside triphosphates, and exogenously added N4 phage DNA; other DNAs are used inefficiently or not at all. Ribonucleoside triphosphates are not required, although they stimulate DNA synthesis. In vitro replication starts at the ends of the N4 genome and moves progressively inward. Initiation occurs through hairpin priming at the 3' end of the genome, but shows a strong preference for the right end. Three N4 gene products (dnp, dbp, and exo) required in vivo for N4 DNA synthesis are absolutely required in the in vitro system. These findings are discussed with respect to the mode of N4 DNA replication.

AB - An in vitro DNA replication system for bacteriophage N4-infected Escherichia coli has been developed. It requires MgCl2, all four deoxyribonucleoside triphosphates, and exogenously added N4 phage DNA; other DNAs are used inefficiently or not at all. Ribonucleoside triphosphates are not required, although they stimulate DNA synthesis. In vitro replication starts at the ends of the N4 genome and moves progressively inward. Initiation occurs through hairpin priming at the 3' end of the genome, but shows a strong preference for the right end. Three N4 gene products (dnp, dbp, and exo) required in vivo for N4 DNA synthesis are absolutely required in the in vitro system. These findings are discussed with respect to the mode of N4 DNA replication.

UR - http://www.scopus.com/inward/record.url?scp=0022975386&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022975386&partnerID=8YFLogxK

M3 - Article

C2 - 3525547

AN - SCOPUS:0022975386

VL - 261

SP - 10506

EP - 10510

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 23

ER -