Fate of exogenous recombinant plasmids introduced into mouse and human cells

Giuseppe Biamonti, Giuliano Della Valle, Daniela Talarico, Fabio Cobianchi, Silvano Riva, Arturo Falaschi

Research output: Contribution to journalArticle

Abstract

We have constructed a number of plasmids selectable in both E. coli and mouse or human cells. Human DNA sequences were inserted and the recombinant plasmids were used to transfect either mouse or human cells by the Ca-phosphate precipitation technique. We have observed that: (i) competent cells uptake large amounts of plasmid DNA; (ii) input plasmids persist in trasformed mammalian cells as free unreplicating circular molecules for up to 20 generations; such persistence does not depend on the presence of selective markers; (iii) plasmids incorporated into mouse L-cells undergo widespread rearrangements (in the absence of replication) entailing mostly deletions of both human and bacterial sequences which yield smaller products; the latter appear to be more stable in a subsequent transformation cycle. Surprisingly such rearrangements are almost totally absent in transformed human KB-cells. This property of human KB-cells mayprove useful for the development of a vector apt at cloning and expressing human DNA sequences. Unlike what has been observed in yeast, no "autonomously replicating sequence" can be detected in mammalian cells by randomly cloning human DNA sequences into a selectable plasmid and screening for an increased transformation efficiency.

Original languageEnglish
Pages (from-to)5545-5561
Number of pages17
JournalNucleic Acids Research
Volume13
Issue number15
DOIs
Publication statusPublished - Aug 12 1985

Fingerprint

Mouse
DNA sequences
Plasmids
Cells
Cloning
Cell
KB Cells
DNA Sequence
Organism Cloning
Rearrangement
Yeast
Escherichia coli
Screening
Phosphates
DNA
Molecules
Human
Recombinant
Yeasts
Phosphate

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Fate of exogenous recombinant plasmids introduced into mouse and human cells. / Biamonti, Giuseppe; Valle, Giuliano Della; Talarico, Daniela; Cobianchi, Fabio; Riva, Silvano; Falaschi, Arturo.

In: Nucleic Acids Research, Vol. 13, No. 15, 12.08.1985, p. 5545-5561.

Research output: Contribution to journalArticle

Biamonti, G, Valle, GD, Talarico, D, Cobianchi, F, Riva, S & Falaschi, A 1985, 'Fate of exogenous recombinant plasmids introduced into mouse and human cells', Nucleic Acids Research, vol. 13, no. 15, pp. 5545-5561. https://doi.org/10.1093/nar/13.15.5545
Biamonti, Giuseppe ; Valle, Giuliano Della ; Talarico, Daniela ; Cobianchi, Fabio ; Riva, Silvano ; Falaschi, Arturo. / Fate of exogenous recombinant plasmids introduced into mouse and human cells. In: Nucleic Acids Research. 1985 ; Vol. 13, No. 15. pp. 5545-5561.
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