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.
Fang
Jia
J
Shanghai Research Center of Biotechnology, the Chinese Academy of Sciences, Shanghai 200233, China. cqchen@server.shcnc.ac.cn
Zhu
Bei
B
Wang
De-Bao
DB
Chen
Chang-Qing
CQ
Duch
Mogens Ryttergaard
MR
Pedersen
Finn Skou
FS