Article

Expression and localization of recombinant human B2 receptors in the methylotrophic yeast Pichia pastoris.

State Key Laboratory of Genetic Engineering, Institute of Biodiversity Science, School of Life Sciences, Fudan University, Shanghai, China.
Genetika (impact factor: 0.44). 08/2006; 42(7):893-7. pp.893-7
Source: PubMed

ABSTRACT The human bradykinin B2 receptor (B2R) fused with green fluorescent protein (GFP) at the C-terminal has been expressed in the methylotrophic yeast of Pichia pastoris. In the expression vector, B2R gene was drove under the highly inducible promoter of alcohol oxidase 1 gene of P. pastoris. By fluorescence activated cell sorting (FACS) analysis and western blot analysis, it was proved that B2R recombinant receptor proteins were expressed at high level in the yeast. Further more, the transformants of P. pastoris were monitored with confocal microscopy, a strong green fluorescence was checked out. The recombinant B2R receptor proteins were mainly located on the plasma membrane proved by immunofluorescence microscopy.

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Keywords

alcohol oxidase 1 gene
 
B2R gene
 
B2R recombinant receptor proteins
 
confocal microscopy
 
FACS
 
fluorescence activated cell sorting
 
human bradykinin B2 receptor
 
immunofluorescence microscopy
 
inducible promoter
 
P. pastoris
 
Pichia pastoris
 
recombinant B2R receptor proteins
 
western blot analysis
 

G X Yang