Crystallization and preliminary X‐ray diffraction analysis of the homing endonuclease I‐CvuI from Chlorella vulgaris in complex with its target DNA. Issue 2 (1st February 2014)

• Record Type: Journal Article
• Authors: Redondo, Pilar; Merino, Nekane; Villate, Maider; Blanco, Francisco J.; Montoya, Guillermo; Molina, Rafael
• Is Part Of: Acta crystallographica. Volume 70:Issue 2(2014:Feb.)
• Page Start: 256
• Page End: 259
• Publication Date: 2014-02-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X1400065X
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Homing endonucleases are highly specific DNA‐cleaving enzymes that recognize long stretches of DNA. The engineering of these enzymes provides novel instruments for genome modification in a wide range of fields, including gene targeting, by inducing specific double‐strand breaks. I‐CvuI is a homing endonuclease from the green alga <italic>Chlorella vulgaris</italic>. This enzyme was purified after overexpression in <italic>Escherichia coli</italic>. Crystallization experiments of I‐CvuI in complex with its DNA target in the presence of Mg²⁺ yielded crystals suitable for X‐ray diffraction analysis. The crystals belonged to the orthorhombic space group <italic>P</italic>2₁2₁2₁, with unit‐cell parameters <italic>a</italic> = 62.83, <italic>b</italic> = 83.56, <italic>c</italic> = 94.40 Å. The self‐rotation function and the Matthews coefficient suggested the presence of one protein–DNA complex per asymmetric unit. The crystals diffracted to a resolution limit of 1.9 Å using synchrotron radiation.</p> </abstract>