Skip Navigation

This Article
Right arrow Print PDF (644K)
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 (17)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Rohan, R. M.
Right arrow Articles by Frels, W. I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rohan, R. M.
Right arrow Articles by Frels, W. I.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1990, Vol. 18, No. 20 6089-6095
© 1990

GENOME STRUCTURE AND MAPPING

Direct sequencing of PCR-amplified junction fragments from tandemly repeated transgenes

Richard M. Rohan+, Donna King*,§ and William I. Frels

US Department of Agriculture, Agricultural Research Service, Reproduction Laboratory, Beltsville Agricultural Research Center Beltsville, MD 20705, USA

*To whom correspondence should be addressed

Received June 4, 1990. Revised September 4, 1990. Accepted September 4, 1990.

When microinjected foreign genes integrate into the genomes of mice, multiple copies are frequently found clustered together at one location. How they concatamerize—by the integration of large linearized concatamers that are formed by simple end-to-end linkage, by circularization of individual DNA fragments and recombination, or by some other means—is not understood. In the transgenic animals studied thus far by ourselves and others, integration frequency and transgene copy number do not seem to be significantly influenced by the complementarity of the ends of the DNA fragments that have been microinjected. We have utilized PCR amplification and DNA sequence analysis to study selected transgene junctions at the nucleotide level. In two transgenic mice carrying the synthetic RSVcat gene (Injected with noncomplementary overhangs on the fragment ends), ends were ‘nibbled’ from 1 to 62 bases before being joined to an adjacent gene copy. Repeated dinucleotides, providing the most minimal of homologies, are present in half of the characterized junctions. Determination of the relative copy number of the junctions in each mouse supports the idea that transgene complexes can undergo additional rearrangements after the initial formation event.

+Present addresses:Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology, School of Medicine, University of Maryland at Baltimore 655 W.Baltimore St., Baltimore, MD 21201

§Department of Pharmacology and Molecular Biology,The Chicago Medical School, 3333 Green Bay Rd., North Chicago, IL 60064-3095, USA


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
 Genome Res.Home page
I S Bevan, R Rapley, and M R Walker
Sequencing of PCR-amplified DNA.
Genome Res., May 1, 1992; 1(4): 222 - 228.
[Abstract] [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.