Construction of His6-tagging vectors allowing single-step purification of GroES and other polypeptides produced in Bacillus subtilis.
Two plasmid expression vectors for Bacillus subtilis have been constructed that direct the synthesis of fusion proteins containing a stretch of six histidine residues (His6) at either the N or C terminus. The His6 tag allows the rapid enrichment of proteins by metal chelate affinity chromatography in a denatured or native state. As an example of the general utility of one of these expression vectors, we produced the GroES protein encoded by the groESL operon of B. stearothermophilus.
Schön
U
U
Lehrstuhl für Genetik, Universität Bayreuth, Germany.
Schumann
W
W
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D018835:Chaperonin 10; D002846:Chromatography, Affinity; D005822:Genetic Vectors; D001411:Geobacillus stearothermophilus / Q000235:genetics; D006360:Heat-Shock Proteins / Q000302:isolation & purification*; D006639:Histidine; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins
D018835:Chaperonin 10; D006360:Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; D006639:Histidine