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Expression and localization of recombinant human EDG-1 receptors in Pichia pastoris |
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| Authors: | Guangxiao Yang Tianlei Liu Wulin Peng Xiaobo Sun Han Zhang Chaoqun Wu Daleng Shen |
| Institution: | (1) Institute of Genetics, State Key Laboratory of Genetic Engineering, Research Centre of Gene Diversity and Designed Agriculture, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Science, Fudan University, Shanghai, 200433, P.R. China;(2) Naval Medical Research Institute, Shanghai, 200433, P.R. China |
| Abstract: | The receptor for human endothelial differentiation gene-1 protein (EDG-1) was C-terminally tagged with green fluorescent protein and expressed in the methylotrophic yeast, Pichia pastoris. EDG-1 expression was driven by the highly inducible alcohol oxidase 1 promoter. Expression of EDG-1 recombinant protein was detected by Western blot analysis and confocal microscopy. The recombinant EDG-1 receptor protein was located in the plasma membrane. Radioligand binding assays demonstrated that the␣EDG-1 receptors expressed in Pichia pastoris␣have specific and saturation binding of 32P-labeled sphingosine 1-phosphate. |
| Keywords: | Endothelial differentiation gene Green fluorescent protein Heterologous expression Pichia pastoris Sphingosine 1-phosphate |
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