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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