Skip Navigation

This Article
Right arrow Print PDF (502K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (41)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Coniey, E. C.
Right arrow Articles by Saunders, J. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Coniey, E. C.
Right arrow Articles by Saunders, J. R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1986, Vol. 14, No. 22 8905-8917
© 1986

Articles

Deletion and rearrangement of plasmld DNA during transformation of Escherichia coli with linear plasmid molecules

Edward C. Coniey1,+, Venetia A. Saunders2 and Jon R. Saunders3

1Departments of Chemistry/Biochemistry Liverpool L3 3AF, UK 2Departments of Biology, Liverpool Polytechnic Liverpool L3 3AF, UK 3Department of Microbiology, University of Liverpool P0 Box 147, Liverpool L69 3BX, UK

Received August 7, 1986. Revised October 23, 1986. Accepted October 23, 1986.

When E.coli was transformed with linearized pBR322 DNA, many transforinants contained recircularized plasmids bearing deletions and other rearrangements. Most aberrant molecuies were <monomeric length and had lost the restriction site used for linearization, with the deleted region extending mono— (type Ia) or bi-directionally (type Ib). Type II deletants were >monomeric but <dimeric and contained the pBR322 sequence in direct repeat with deletion at one or both junctions (type IIa) or in inverted repeat with loss of sequence at both junctions (type IIb). Type III deletants were >dimeric but <trimeric, consisting of pBR322 sequences in both direct and inverse repeat with deletions at two or more junctions. Transformation frequencies for linear DNA were drastically reduced in xth-1– bacteria with type hib deletants predominating in transformants. This indicates that exonuclease III is important for perfect recyclization of plasmids and the generation of type I deletants. In vivo recyclization of in vitro ligation products explains many of the aberrant DNA molecules that are encountered during gene cloning.

+Present address: Enzymology of Genetic Recombination Laboratory, Imperial Cancer Research Fund, Clare Hall Laboratories, Potters Bar, Heits EN6 3LD, UK


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
B. Kosmider and R. D. Wells
Double-strand breaks in the myotonic dystrophy type 1 and the fragile X syndrome triplet repeat sequences induce different types of mutations in DNA flanking sequences in Escherichia coli
Nucleic Acids Res., November 14, 2006; 34(19): 5369 - 5382.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
A. C. Adam, G. González-Blasco, M. Rubio-Texeira, and J. Polaina
Transformation of Escherichia coli with DNA from Saccharomyces cerevisiae Cell Lysates
Appl. Envir. Microbiol., December 1, 1999; 65(12): 5303 - 5306.
[Abstract] [Full Text]

Home page
 Genome ResHome page
D H Jones
PCR mutagenesis and recombination in vivo.
Genome Res., June 1, 1994; 3(6): S141 - S148.
[PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.