Overexpression, purification and crystallization of BamHI endonuclease.
The type II restriction endonuclease BamHI has been expressed in E. coli, producing 100-fold more enzyme than the wild type Bacillus amyloliquefaciens H strain. This high yield has facilitated purification to homogeneity of large amounts of the enzyme, along with its crystallization in a form which diffracts to at least 1.9 A in X-ray analysis.
Jack
W E
WE
New England Biolabs, Inc., Beverly, MA 01915.
Greenough
L
L
Dorner
L F
LF
Xu
S Y
SY
Strzelecka
T
T
Aggarwal
A K
AK
Schildkraut
I
I
D001407:Bacillus / Q000201:enzymology; D003001:Cloning, Molecular; D003460:Crystallization; D015245:Deoxyribonuclease BamHI / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D015245:Deoxyribonuclease BamHI