[Comparison of two transmemembrane proteins as fusion partner for protein expression on the surface of cell].
The expression of a soluble protein on cell surface is often desirable for study of a functional protein, wide application of a protein or investigation of protein-protein interaction. The expression of a soluble protein on the surface of a cell is often achieved by genetically linking a protein to the extra-cellular fragment of a transmembrane partner. In this study, the myc epitope was linked with N terminal of transmembrane proteins either A2TM or deltaLNGFR amplified by overlapping PCR. The plasmids expressing fusion protein were transfected into 293FT cells and the expression of target proteins was evaluated by fluorescent microscope, flow cytometry and Western blotting. The results of flow cytometry revealed that both A2TM and deltaLNGFR were expressed on the cell surface, but A2TM could only be detected with high copy number. Western blotting showed that the expression level of deltaLNGFR was very high and protein was heavily glycosylated, by contrast the expression of A2TM was hardly detected. The results indicate that glycosylated deltaLNGFR is a good candidate partner for the expression of a soluble protein on the cell surface.
Liu
Qingjun
Q
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Han
Huamin
H
Zhang
Zhaoshan
Z
Gao
Bin
B
D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism*; D005802:Genes, MHC Class II / Q000235:genetics; D015789:HLA-A2 Antigen / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D050576:Membrane Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*
D051017:Apoptosis Regulatory Proteins; C410727:BEX3 protein, human; D015789:HLA-A2 Antigen; D050576:Membrane Fusion Proteins; D008565:Membrane Proteins; D016271:Proto-Oncogene Proteins c-myc