title: PCR-directed preparation and single-step purification of highly active histidine-tagged restriction endonuclease HgiBI (GGWCC).
abstract: 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.
authors.affiliation: Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Blum
authors.forename: G
authors.initials: G
authors.lastname: Horst
authors.forename: M
authors.initials: M
authors.lastname: Kröger
mesh_terms: 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
chemical_list: D011993:Recombinant Fusion Proteins; D006639:Histidine; D015252:Deoxyribonucleases, Type II Site-Specific; C038308:GGWCC-specific type II deoxyribonucleases