title: Production of biologically active recombinant avidin in baculovirus-infected insect cells.
abstract: An efficient lepidopteran insect cell system was established for the expression of a recombinant form of chicken egg-white avidin. The gene product was obtained in both secreted and intracellular forms, and biologically active recombinant avidin was isolated using affinity chromatography on an iminobiotin-agarose column. Similar to the known quaternary structure of the native egg-white protein, the purified recombinant protein was glycosylated and assembled mainly into tetramers. Like native avidin, the recombinant tetramer also exhibited a high level of thermostability, and was further stabilized upon binding biotin. The biotin-binding and structural properties of the recombinant avidin are thus similar to those of the natural egg-white protein, and the insect system is appropriate both for future site-directed mutagenesis studies and for the production of avidin fusion proteins.
authors.affiliation: Department of Biological and Environmental Science, University of Jyväskylä, Finland.
authors.forename: K J
authors.initials: KJ
authors.lastname: Airenne
authors.forename: C
authors.initials: C
authors.lastname: Oker-Blom
authors.forename: V S
authors.initials: VS
authors.lastname: Marjomäki
authors.forename: E A
authors.initials: EA
authors.lastname: Bayer
authors.forename: M
authors.initials: M
authors.lastname: Wilchek
authors.forename: M S
authors.initials: MS
authors.lastname: Kulomaa
mesh_terms: D000818:Animals; D001360:Avidin / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D016367:Baculoviridae / Q000235:genetics; D001710:Biotin / Q000031:analogs & derivatives; D002645:Chickens / Q000235:genetics; D002846:Chromatography, Affinity / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006031:Glycosylation; D011487:Protein Conformation; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012685:Sepharose; D018411:Spodoptera / Q000166:cytology / Q000821:virology
chemical_list: D011994:Recombinant Proteins; D001360:Avidin; D001710:Biotin; D012685:Sepharose; C028072:2-iminobiotin