Periplasmic targeting of recombinant proteins in Escherichia coli.
Targeting recombinant protein production to the periplasmic space of Escherichia coli presents several advantages over cytoplasmic production in inclusion bodies and at the same time overcomes the low productivity problem often associated with culture medium secretion. This chapter presents a strategy for periplasmic production of recombinant proteins fused to synthetic Z domains derived from staphylococcal protein A. Expression, purification, and monitoring strategies are discussed using green fluorescent protein and human proinsulin as model proteins.
Mergulhão
Filipe J
FJ
LEPAE, Faculty of Engineering of the University of Porto, Chemical Engineering Department, Porto, Portugal.
Monteiro
Gabriel A
GA
D003470:Culture Media / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D019897:Periplasm / Q000378:metabolism*; D011384:Proinsulin / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
D003470:Culture Media; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011384:Proinsulin