Mechanism of DNA Recognition by the Restriction Enzyme EcoRV

Role of thymidine residues in DNA recognition by the EcoRI and EcoRV restriction endonucleases.

We have synthesized a series of oligonucleotides containing the EcoRI (GAATTC) or EcoRV (GATATC) recognition site within which or adjacent to which thymidine was substituted by uridine or derivatives of uridine. The effects of these substitutions on the rate of the EcoRI and EcoRV catalyzed cleavage reaction were investigated. Our results show that most of the substitutions within the site are ...

Analysis of the recognition mechanism involved in the EcoRV catalyzed cleavage of DNA using modified oligodeoxynucleotides.

We have prepared a series of undecadeoxynucleotides that contain changes in the functional group pattern present within the EcoRV recognition site - GATATC-. Oligonucleotides were synthesized on solid phase using normal and modified beta-cyanoethylphosphoramidites and analyzed in steady state cleavage experiments with the EcoRV restriction endonuclease. The following groups appear to interact s...

Characterization of Avian Pasteurella multocida Isolates by Protein Profiles and Restriction Enzyme Analysis of Chromosomal DNA

The average spacing of restriction enzyme recognition sites in DNA.

The discovery of naturally occurring enzymes which cleave DNA at sites specific to particular nucleotide sequences has had a great impact on molecular biology. The function of these enzymes in uivo is to protect bacterial cells from viral invasion by degradation of foreign DNA. Several hundred of these “restriction” enzymes are known and they are a very common tool both for analysis and manipul...

Fluorescent substrates for the EcoRV restriction endonuclease.

In the presence of Mg2+, the EcoRV restriction endonuclease cleaves DNA specifically at the sequence GAT'ATC (where denotes the point of scission) [ I ] . Even the change of a single base pair within this sequence will lead to a million fold reduction of EcoRV activity [2]. Paradoxically, gel retardation experiments in the absence of Mg2+ show that, unlike EcoRl and many other type 11 restricti...