Massively parallel characterization of restriction endonucleases

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Massively parallel characterization of restriction endonucleases

Restriction endonucleases are highly specific in recognizing the particular DNA sequence they act on. However, their activity is affected by sequence context, enzyme concentration and buffer composition. Changes in these factors may lead to either ineffective cleavage at the cognate restriction site or relaxed specificity allowing cleavage of degenerate 'star' sites. Additionally, uncharacteriz...

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The characterization of restriction endonucleases: the work of Hamilton Smith.

Hamilton Othanel Smith was born in 1931 in New York City. In 1937, he and his family moved to Champaign-Urbana, Illinois, because his father had joined the faculty of the department of education at the University of Illinois. As a boy, Smith was interested in chemistry, electricity, and electronics, and he spent many hours with his brother in their basement laboratory, which was stocked with su...

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Characterization of the 5-hydroxymethylcytosine-specific DNA restriction endonucleases

In T4 bacteriophage, 5-hydroxymethylcytosine (5hmC) is incorporated into DNA during replication. In response, bacteria may have developed modification-dependent type IV restriction enzymes to defend the cell from T4-like infection. PvuRts1I was the first identified restriction enzyme to exhibit specificity toward hmC over 5-methylcytosine (5mC) and cytosine. By using PvuRts1I as the original me...

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The type IIB restriction endonucleases.

The endonucleases from the Type IIB restriction-modification systems differ from all other restriction enzymes. The Type IIB enzymes cleave both DNA strands at specified locations distant from their recognition sequences, like Type IIS nucleases, but they are unique in that they do so on both sides of the site, to liberate the site from the remainder of the DNA on a short duplex. The fact that ...

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 2013

ISSN: 1362-4962,0305-1048

DOI: 10.1093/nar/gkt257