Cell surface display of RAGE variable domain on Escherichia coli for establishment of affinity binding assay. The 11th International Congress ASEAN Food Conference

in International Symposium (oral presentation paper), 國際研討會(全文口頭發表)
標題Cell surface display of RAGE variable domain on Escherichia coli for establishment of affinity binding assay. The 11th International Congress ASEAN Food Conference
出版類型國際研討會(全文口頭發表)
出版年度2009
AuthorsMei-Li Wu, 吳美莉
其他編號0000
中文摘要

Abstract
The receptor for advanced glycation end products (RAGE) belongs to the immunoglobulin superfamily of cell surface receptors. Activation of RAGE by AGEs may lead to proinflammatory signaling, likely to accelerate the processes of coronary atherosclerotic development, especially in diabetic patients. The ice nucleation protein (INP) is a membrane-associated protein from Pseudomonas syringae, which targeted foreign proteins on the cell surface. In this study, RAGE V-type domain (RAGE-V) cDNA sequence have been cloned using reverse transcription-polymerase chain reaction (RT-PCR) from Human promyelocytic leukemia (HL-60). We constructed a cell surface display system by using INPN as an anchor motif for surface expression of RAGE-V on E. coli. Western blot analysis identified the synthesis of INPN-RAGE-V fusion protein of the expected size. Immunofluorescence microscopy further confirmed that RAGE-V was anchored on the cell surface. Time course for the growth profile clearly demonstrated that the INP integration did not significantly affect cell growth under IPTG induction. Whole-cell enzyme-linked immunosorbent assay confirmed that the displayed RAGE-V showed binding activity toward AGEs. The expressed RAGE-V can be applied as an in vitro screening system for screening RAGE blocking constituent from herbs.
Key words: advanced glycation end products; RAGE; variable domain; surface display

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