[Eukaryotic expression of human LIGHT-Fc chimeric molecule and its anti-tumor activity]

Construct eukaryotic expression plasmid of hLIGHT-Fc gene and express hLIGHT-Fc fusion protein with high biological activity.

Utilizing PCR to clone the human LIGHT cDNA from a normal human activated T cells cDNA library phAD. CAD, then using SOE (splicing by overlap extension) technique to fuse CD137 signal peptide gene into LIGHT extramembrane domain encoding sequence, inserting this recombinated LIGHT cDNA with human IgGI Fc cDNA into the eukaryotic expression plasmid pcDNA3, expressed it transiently in 293T cells, and purified by recombinated protein A affinity chromatography column, at last we checked its Mr, purity and anti-tumor activity.

LIGHT-Fc gene's ORF is right and coincident with we expected, then its expression was confirmed by ELISA and SDS-PAGE, and we proved that the purified LIGHT-Fc protein could inhibit the growth of some cultured tumor cell lines by MTT.

we got a purified recombinated protein of LIGHT-Fc with anti-tumor activity and the anticipated Mr, thus lay a foundation for further works on LIGHT such as its role in immune homeostasis,the mechanism of its receptors' signal transduction, et al.