Restriction site

The restriction site (synonym restriction sequence ) is a DNA sequence to which a certain restriction enzyme binds and cuts the DNA.

properties

Each restriction enzyme has a characteristic DNA sequence (recognition sequence) of mostly three to eight base pairs to which it can bind. The two different restriction sites and the restriction enzymes cutting at this point are selected in the course of cloning so that, after restriction digestion of the DNA, ligation with the vector can take place in the correct orientation. Since restriction enzymes mostly bind to the recognition sequence as homodimers , the recognition sequences are palindromic . Isoschizomers produce the same DNA cut, while neoschizomers only have the same recognition sequence .

Some restriction sites produce blunt ends (interfaces without a base overhang), others sticky ends (interfaces with a base overhang). A base overhang allows the binding of another DNA with a complementary base overhang by base pairing , which makes ligation by a DNA ligase much more efficient.

REBASE is an online database that lists restriction sites.

literature

• Cornel Mülhardt: The Experimenter: Molecular Biology / Genomics. Sixth edition. Spectrum Akademischer Verlag, Heidelberg 2008, ISBN 3-8274-2036-9 .
• J. Sambrook , T. Maniatis , DW Russel: Molecular cloning: a laboratory manual. 3rd edition (2001), Cold Spring Harbor Laboratory Press. ISBN 0-87969-577-3 .

Individual evidence

1. ↑ Peter J. Russell: iGenetics: A Mendelian Approach . Benjamin Cummings, 2006, ISBN 978-0805346664 .
2. ↑ Albert L. Lehninger, David L. Nelson, Michael M. Cox: Principles of Biochemistry , 5th. Edition, WH Freeman and Company, New York, NY 2008, ISBN 978-0-7167-7108-1 . P. 305.
3. ↑ Mohammad Mousavi-Khattat, Adele Rafati, Pooria Gill: Fabrication of DNA nanotubes using origami-based nanostructures with sticky ends. In: Journal of Nanostructure in Chemistry. 5, 2015, p. 177, doi : 10.1007 / s40097-015-0148-z .
4. ^ S. Gao, J. Zhang, T. Miao, D. Ma, Y. Su, Y. An, Q. Zhang: A simple and convenient sticky / blunt-end ligation method for fusion gene construction. In: Biochemical genetics. Volume 53, Number 1–3, April 2015, pp. 42–48, doi : 10.1007 / s10528-015-9669-x , PMID 25820211 .
5. ↑ RJ Roberts, T. Vincze, J. Posfai, D. Macelis: REBASE - a database for DNA restriction and modification: enzymes, genes and genomes. In: Nucleic acids research. Volume 38, Database issue January 2010, pp. D234-D236, doi : 10.1093 / nar / gkp874 , PMID 19846593 , PMC 2808884 (free full text).
6. ↑ RJ Roberts, T. Vincze, J. Posfai, D. Macelis: REBASE - a database for DNA restriction and modification: enzymes, genes and genomes. In: Nucleic acids research. Volume 43, Database issue January 2015, pp. D298 – D299, doi : 10.1093 / nar / gku1046 , PMID 25378308 , PMC 4383893 (free full text).