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Biotechnol Lett ; 31(1): 49-55, 2009 Jan.
Article in English | MEDLINE | ID: mdl-18797996

ABSTRACT

The three beta adrenergic receptor subtypes, beta1-, beta2- and beta3-, were expressed in the methylotrophic yeast Pichia pastoris. These receptors were N-terminally fused to the enhanced green fluorescent protein (EGFP) and the fluorescent properties of EGFP were used: (1) to select the recombinant strains, (2) to monitor the expression of the fluorescent receptors, and (3) to monitor the purification of the receptors by immobilized metal affinity chromatography. We demonstrate here that Pichia pastoris can be an alternative host to express and purify milligram amounts of human beta adrenergic receptors.


Subject(s)
Green Fluorescent Proteins/metabolism , Pichia/metabolism , Receptors, Adrenergic, beta/isolation & purification , Receptors, Adrenergic, beta/metabolism , Fluorescence , Green Fluorescent Proteins/genetics , Humans , Immunoblotting , Kinetics , Receptors, Adrenergic, beta/genetics , Receptors, Adrenergic, beta-1/genetics , Receptors, Adrenergic, beta-1/isolation & purification , Receptors, Adrenergic, beta-1/metabolism , Receptors, Adrenergic, beta-2/genetics , Receptors, Adrenergic, beta-2/isolation & purification , Receptors, Adrenergic, beta-2/metabolism , Receptors, Adrenergic, beta-3/genetics , Receptors, Adrenergic, beta-3/isolation & purification , Receptors, Adrenergic, beta-3/metabolism , Silver Nitrate , Solubility
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