含识别位点DNA的限制性内切酶PvuII的表达、纯化及结晶

Expression, purification, and crystallization of restriction endonuclease PvuII with DNA containing its recognition site.

作者信息

Balendiran K, Bonventre J, Knott R, Jack W, Benner J, Schildkraut I, Anderson J E

机构信息

W. M. Keck Structural Biology Laboratory, Cold Spring Harbor Laboratory, New York 11724.

出版信息

Proteins. 1994 May;19(1):77-9. doi: 10.1002/prot.340190110.

DOI:10.1002/prot.340190110

PMID:8066089

Abstract

We have overexpressed the type II restriction endonuclease PvuII (R-PvuII) in E. coli, prepared large amounts of the homogeneous enzyme, and crystallized it with an oligonucleotide carrying a PvuII recognition site. The cocrystals are orthorhombic space group P2(1)2(1)2(1) with cell constants a = 95.8 A, b = 86.3 A, c = 48.5 A, and diffract X-rays to at least 2.7 A. There is a complex of two protein subunits and one oligonucleotide duplex in the asymmetric unit.

摘要

我们在大肠杆菌中过表达了II型限制性内切酶PvuII（R-PvuII），制备了大量的纯酶，并使其与携带PvuII识别位点的寡核苷酸结晶。共晶体属于正交空间群P2(1)2(1)2(1)，晶胞参数a = 95.8 Å，b = 86.3 Å，c = 48.5 Å，X射线衍射分辨率至少为2.7 Å。不对称单元中有两个蛋白质亚基和一个寡核苷酸双链体的复合物。