Cleaving DNA at Any Predetermined Site with Adapter-Primers and Class-IIS Restriction Enzymes
- 22 April 1988
- journal article
- research article
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 240 (4851), 504-506
- https://doi.org/10.1126/science.2833816
Abstract
A four-component system has been designed that makes it possible to prepare a double-stranded (ds) DNA fragment; one fragment end is predesigned (by the use of a class-IIS restriction enzyme and adapter-primer), and the other end corresponds to any normal restriction cut. The system is composed of the phage M13mp7 single-stranded (ss) target DNA; the Fok I restriction enzyme; an oligodeoxynucleotide adapter-primer, which permits one to introduce Fok I cuts at any specified site in the target DNA; and DNA polymerase, which converts the ss target into a ds form ready for cloning. In this system, the oligodeoxynucleotide adapter-primer serves several purposes. The 5' hairpin ds domain of the adapter-primer contains the Fok I recognition site. Its 3' ss domain selects a complementary site on the target ss DNA, hybridizes with it to form the ds cleavage site, and serves as a primer to convert the ss M13mp7 target to ds DNA.This publication has 15 references indexed in Scilit:
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