Skip Navigation

This Article
Right arrow Print PDF (4962K)
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 (42)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Demidov, V. V.
Right arrow Articles by Nielsen, P. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Demidov, V. V.
Right arrow Articles by Nielsen, P. E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1994, Vol. 22, No. 24 5218-5222
© 1994

Articles

Electron microscopy mapping of oligopurine tracts in duplex DNA by peptide nucleic acid targeting

Vadim V. Demidov1,2,3, Dmitry I. Cherny1, Alexey V. Kurakin1, Michael V. Yavnilovich1,+, Vladislav A. Malkov1,§, Maxim D. Frank-Kamenetskii2, Spren H. Sdnnichsen3 and Peter E. Nielsen3,*

1Institute of Molecular Genetics, Russian Academy of Sciences 46 Kurchatov Square, Moscow 123182, Russia 2Center for Advanced Biotechnology, Department of Biomedical Engineering, Boston University 36 Cummington Street, Boston, MA 02215, USA 3Center for Biomolecular Recognition, IMBG Department B, The Panum Institute Blegdamsvej 3c, DK-2200 Copenhagen N, Denmark

+Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel

§Genetics and Biochemistry Branch, National Institute of Diabetes and Kidney and Digestive Diseases, NTH, Bethesda, MD 20892, USA

*To whom correspondence should be addressed

Received September 24, 1994. Accepted November 9, 1994.

Biotinylated homopyrlmidlne decamer peptide nucleic acids (PNAs) are shown to form sequence-specific and stable complexes with complementary oligopurine targets in linear double-stranded DNA. The non-covalent complexes are visualized by electron microscopy (EM) without chemical fixation using streptavldln as an EM marker. The triplex stoichiometry of the PNA-DNA complexes (two PNA molecules presumably binding by Watson - Crick and Hoogsteen pairing with one of the strands of the duplex DNA) is indicated by the appearance of two streptavldin ‘beads’ per target site In some micrographs, and is also supported by the formation of two retardation bands in a gel shift assay. Quantitative analysis of the positions of the streptavldln ‘beads’ revealed that under optimized conditions PNA-DNA complexes are preferably formed with the fully complementary target. An increase In either the PNA concentration or the incubation time leads to binding at sites containing one or two mismatches. Our results demonstrate that biotinylated PNAs can be used for EM mapping of short targets in duplex DNA.


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
 Biophys. JHome page
A. Abibi, E. Protozanova, V. V. Demidov, and M. D. Frank-Kamenetskii
Specific versus Nonspecific Binding of Cationic PNAs to Duplex DNA
Biophys. J., May 1, 2004; 86(5): 3070 - 3078.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. C. Boffa, P. L. Morris, E. M. Carpaneto, M. Louissaint, and V. G. Allfrey
Invasion of the CAG Triplet Repeats by a Complementary Peptide Nucleic Acid Inhibits Transcription of the Androgen Receptor and TATA-binding Protein Genes and Correlates with Refolding of an Active Nucleosome Containing a Unique AR Gene Sequence
J. Biol. Chem., May 31, 1996; 271(22): 13228 - 13233.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Bouziane, D. I. Cherny, J.-F.ço. Mouscadet, and C. Auclair
Alternate Strand DNA Triple Helix-mediated Inhibition of HIV-1 U5 Long Terminal Repeat Integration in Vitro
J. Biol. Chem., April 26, 1996; 271(17): 10359 - 10364.
[Abstract] [Full Text] [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.