Expression and purification of thioredoxin fusion proteins.
This unit describes a gene fusion expression system that uses thioredoxin, the product of the Escherichia coli trxA gene, as the fusion partner. The system is particularly useful for high-level production of soluble fusion proteins in the E. coli cytoplasm; in many cases heterologous proteins produced as thioredoxin fusion proteins are correctly folded and display full biological activity. Protein fusions to His-patch Trx can usually be purified in a single step from cell lysates. Additional protocols describe E. coli cell lysis using a French pressure cell and fractionation, osmotic release of thioredoxin fusion proteins from the E. coli cytoplasm, and heat treatment to purify some thioredoxin fusion proteins.
McCoy
J
J
Genetics Institute, Cambridge, Massachusetts, USA.
Lavallie
E
E
D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D009997:Osmotic Pressure; D011312:Pressure; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism*
D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; C517609:TrxA protein, E coli; D013879:Thioredoxins