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Isolation of human sequences that replicate autonomously in human cells.

Publication ,  Journal Article
Krysan, PJ; Haase, SB; Calos, MP
Published in: Molecular and cellular biology
March 1989

We have isolated a heterogeneous collection of human genomic sequences which replicate autonomously when introduced into human cells. The novel strategy for the isolation of these sequences involved cloning random human DNA fragments into a defective Epstein-Barr virus vector. This vector alone was unable to replicate in human cells, but appeared to provide for the nuclear retention of linked DNA. The human sequences persist in a long-term replication assay (greater than 2 months) in the presence of the viral nuclear retention sequences. Using a short-term (4-day) assay, we showed that the human sequences are able to replicate in the absence of all viral sequences. The plasmids bearing human sequences were shown to replicate based on the persistence of MboI-sensitive plasmid DNA in the long-term assay and the appearance of DpnI-resistant DNA in the short-term assay. The human sequences were shown to be responsible for the replication activity and may represent authentic human origins of replication.

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Published In

Molecular and cellular biology

DOI

EISSN

1098-5549

ISSN

0270-7306

Publication Date

March 1989

Volume

9

Issue

3

Start / End Page

1026 / 1033

Related Subject Headings

  • Replicon
  • Plasmids
  • Humans
  • Herpesvirus 4, Human
  • Genetic Vectors
  • Genetic Markers
  • Genes
  • Developmental Biology
  • DNA Replication
  • Cloning, Molecular
 

Citation

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Krysan, P. J., Haase, S. B., & Calos, M. P. (1989). Isolation of human sequences that replicate autonomously in human cells. Molecular and Cellular Biology, 9(3), 1026–1033. https://doi.org/10.1128/mcb.9.3.1026-1033.1989
Krysan, P. J., S. B. Haase, and M. P. Calos. “Isolation of human sequences that replicate autonomously in human cells.Molecular and Cellular Biology 9, no. 3 (March 1989): 1026–33. https://doi.org/10.1128/mcb.9.3.1026-1033.1989.
Krysan PJ, Haase SB, Calos MP. Isolation of human sequences that replicate autonomously in human cells. Molecular and cellular biology. 1989 Mar;9(3):1026–33.
Krysan, P. J., et al. “Isolation of human sequences that replicate autonomously in human cells.Molecular and Cellular Biology, vol. 9, no. 3, Mar. 1989, pp. 1026–33. Epmc, doi:10.1128/mcb.9.3.1026-1033.1989.
Krysan PJ, Haase SB, Calos MP. Isolation of human sequences that replicate autonomously in human cells. Molecular and cellular biology. 1989 Mar;9(3):1026–1033.

Published In

Molecular and cellular biology

DOI

EISSN

1098-5549

ISSN

0270-7306

Publication Date

March 1989

Volume

9

Issue

3

Start / End Page

1026 / 1033

Related Subject Headings

  • Replicon
  • Plasmids
  • Humans
  • Herpesvirus 4, Human
  • Genetic Vectors
  • Genetic Markers
  • Genes
  • Developmental Biology
  • DNA Replication
  • Cloning, Molecular