Expression in BHK Cells of Two Human TNF
Receptors and
Their Interaction with hTNF-α
FANG Jia, WANG De-Bao1 and CHEN Chang-Qing*
( Shanghai Research Center of Biotechnology, the Chinese Academy of
Sciences, Shanghai 200233, China；
1Shanghai Institute of Biochemistry, the Chinese Academy of
Sciences, Shanghai 200031, China )
Abstract Two human tumor necrosis
factor receptor genes were cloned into eukaryotic expression vectors pcDNA3,
pDR2 and pXJ-41, respectively. Those vectors were transfected into BHK-21 cells
and the expression of hTNFRs was identified and demonstrated by binding of ［125I］hTNF-α and Scatchard analysis.
The expression of two hTNFRs in BHK cells at the RNA transcriptional and the
protein translation levels were also demonstrated by RT-PCR and indirect ELISA.
However, hTNF-α did not appear cytotoxic to these cells. The competitive
binding activity of mutant hTNF-α-R2K and wild type hTNF-α with these two
receptors, respectively, were assayed. These results laid basis for further
studies on structure-function relationships of these receptors as well as the
hTNF-α.
Key Words hTNF receptors; expression; hTNF-α;
structure and function
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