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[A new site-specific endodeoxyribonuclease EcoHI].

L I Glatman, A A Terekhov, K V Kalnin

Molekuliarnaia Genetika, Mikrobiologiia I Virusologiia

|March 1, 1990

Summary

A novel restriction enzyme, EcoHI, was identified from E. coli. This enzyme recognizes a specific DNA sequence and offers a high yield, making it valuable for molecular biology applications.

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Area of Science:

Molecular Biology
Enzymology
Microbiology

Background:

Site-specific endonucleases are crucial tools in molecular biology.
The discovery of new restriction enzymes expands the toolkit for DNA manipulation.
Escherichia coli is a common host for isolating and producing recombinant proteins and enzymes.

Purpose of the Study:

To isolate and characterize a new type II site-specific endonuclease from Escherichia coli.
To determine the DNA recognition sequence and cleavage site of the novel enzyme.
To assess the enzyme's yield and the characteristics of the producing bacterial strain.

Main Methods:

Isolation and purification of the restriction endonuclease from Escherichia coli.
DNA sequencing and analysis to determine the recognition site.
Enzyme activity assays to quantify yield.
Characterization of the producing strain (Escherichia coli HI).

Main Results:

A new type II site-specific endonuclease, EcoHI, was successfully isolated and characterized.
EcoHI recognizes the DNA sequence 5'-C C (C/G) G G-3' with cleavage between the 4th and 5th nucleotides.
It is an isoshizomer of the restriction endonuclease CauII.
The enzyme yield is 2500 units per gram of biomass.
The producing strain, E. coli HI, is nonpathogenic and possesses antibiotic resistance markers for selective cultivation.

Conclusions:

EcoHI is a novel restriction enzyme with potential applications in molecular biology.
Its specific recognition sequence and high yield make it a valuable addition to existing restriction enzyme libraries.
The characteristics of the producing strain facilitate its large-scale cultivation and enzyme production.

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