PCR-directed preparation and single-step purification of highly active histidine-tagged restriction endonuclease HgiBI (GGWCC).
The polymerase chain reaction was used to produce His6 fusion proteins via deletion of an intervening piece of DNA. The generally applicable method was performed using a standard primer with the advantage that the fusion does not produce additional amino acids. In a single-step purification highly purified, enzymatically active restriction endonuclease was obtained.
Blum
E
E
Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Germany.
Horst
G
G
Kröger
M
M
D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D015252:Deoxyribonucleases, Type II Site-Specific / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D006639:Histidine; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D016324:Sequence Tagged Sites
D011993:Recombinant Fusion Proteins; D006639:Histidine; D015252:Deoxyribonucleases, Type II Site-Specific; C038308:GGWCC-specific type II deoxyribonucleases