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FseI, a new type II restriction endonuclease that recognizes the octanucleotide sequence 5' GGCCGGCC 3'
J M Nelson1, S M Miceli, M P Lechevalier
1Cold Spring Harbor Laboratory, NY 11724.
Nucleic Acids Research
|April 25, 1990
Summary
A novel restriction enzyme, FseI, was purified from Frankia species. This enzyme recognizes a specific DNA sequence and shows potential for genomic applications due to its site frequency similar to NotI.
Area of Science:
- Molecular Biology
- Enzymology
- Genomics
Background:
- Restriction enzymes are crucial tools in molecular biology for DNA manipulation.
- Identifying novel restriction enzymes expands the toolkit for genetic engineering and analysis.
Purpose of the Study:
- To partially purify and characterize a novel Type II restriction endonuclease, FseI, from Frankia species.
- To determine the DNA recognition sequence and cleavage specificities of FseI.
- To compare the genomic site frequency of FseI with existing restriction enzymes.
Main Methods:
- Partial purification of FseI from Frankia species (NRRL 18528).
- DNA cleavage assays using various viral and plasmid DNA substrates.
- Determination of the FseI recognition sequence (5' GGCCGGCC 3').
Main Results:
- FseI was partially purified and identified as a Type II restriction endonuclease.
- FseI specifically cleaves Adenovirus 2 DNA at three sites.
- FseI does not cleave DNA from bacteriophages lambda, T7, phi X174, SV40, pUC18, or pBR322.
- The enzyme recognizes and cleaves the octanucleotide sequence 5' GGCCGGCC 3'.
Conclusions:
- FseI is a novel restriction enzyme with unique specificity.
- The recognition sequence and cleavage pattern of FseI have been elucidated.
- The frequency of FseI sites in the human genome is comparable to that of NotI, suggesting potential utility in large-scale genome analysis.