Fate of exogenous recombinant plasmids introduced into mouse and human cells (original) (raw)

Journal Article

,

Search for other works by this author on:

,

1

Dipartmentimento di Genetica e Microbiologia, Universitá di Pavia

Via S.Epifanio, 14-27100 Pavia, Italy

Search for other works by this author on:

,

1

Dipartmentimento di Genetica e Microbiologia, Universitá di Pavia

Via S.Epifanio, 14-27100 Pavia, Italy

Search for other works by this author on:

,

Search for other works by this author on:

,

Search for other works by this author on:

* To Whom correspondence should be addressed

Search for other works by this author on:

Revision received:

28 June 1985

Published:

12 August 1985

• PDF
• Split View
• • Article contents
  • Figures & tables
  • Video
  • Audio
  • Supplementary Data
• Cite

Cite

Giuseppe Biamonti, Giuliano Della Valle, Daniela Talarico, Fabio Cobianchi, Silvano Riva, Arturo Falaschi, Fate of exogenous recombinant plasmids introduced into mouse and human cells, Nucleic Acids Research, Volume 13, Issue 15, 12 August 1985, Pages 5545–5561, https://doi.org/10.1093/nar/13.15.5545
Close

Navbar Search Filter Mobile Enter search term Search

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.

This content is only available as a PDF.

© IRL Press Limited

I agree to the terms and conditions. You must accept the terms and conditions.

Submit a comment

Name

Affiliations

Comment title

Comment

You have entered an invalid code

Thank you for submitting a comment on this article. Your comment will be reviewed and published at the journal's discretion. Please check for further notifications by email.

Citations

Views

Altmetric

Metrics

Total Views 69

12 Pageviews

57 PDF Downloads

Since 4/1/2017

Citations

34 Web of Science

×

Email alerts

Citing articles via

• Latest

• Most Read

• Most Cited

More from Oxford Academic