Overexpressed Human TNF Receptor-75 Can Independently Mediate hTNFalpha-induced Cytotoxicity in BHK-21 Cells.
A recombinant bicistronic expression vector was constructed containing human TNF receptor-75 gene and encephalomyocarditis virus(EMCV)internal ribosome entry site(IRES), followed by a neomycin phosphotransferase gene as selectable marker. BHK-21 cells were transfected with this vector using electroporation method and positive clones overexpressing the genes of interest were obtained after selection with G-418. The expression of two hTNFRs in those cells at RNA transcriptional and the protein translation levels had been proved by RT-PCR and indirect ELISA. It was found that the hTR75 could mediate cytotoxicity independently in BHK-21 cells after those cells were treated by hTNFalpha. Furthermore, hTR55 did not display the synergistic activity on this function of hTR75. These results put forward a new way for further studies on structure and function relationships of hTR75 and the interactions between two hTNFRs.
Shanghai Research Center of Biotechnology, the Chinese Academy of Sciences, Shanghai 200233, China. cqchen@server.shcnc.ac.cn
Jia
J
Fang
Bei
B
Zhu
De-Bao
DB
Wang
Chang-Qing
CQ
Chen
Mogens Ryttergaard
MR
Duch
Finn Skou
FS
Pedersen