2004•PubMedRequires access

[Bioassay of the soluble human tumor necrosis factor receptor I recombinant plasmid expression in vitro].

Chenrong Xu, Jin-cai Zhang, Chuan-jiang Zhao, Yunhui Zhang

Abstract

OBJECTIVE: To detect the expression of recombinant plasmid PcDNA3.1-sTNFRI in vitro and evaluate the bioactivity of expressed sTNFRI. METHODS: CHO cells were transfected with recombinant plasmid PcDNA3.1-sTNFRI by liposome. sTNFRI in cell culture supernatant was detected by ELISA and sTNFRI blockage of TNF-alpha cytotoxicity in L929 cells evaluated by MTT assay. RESULTS: The expression of sTNFRI in transfected cell culture supernatant was higher than control groups (P < 0.001). The expressed sTNFRI could significantly neutralize TNF-alpha cytotoxicity in L929 cells. CONCLUSIONS: These results showed that the recombinant plasmid PcDNA3.1-sTNFRI can be expressed in mammalian cells and the recombinant sTNFRI has biological function.

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Available abstract

Key concepts: Recombinant DNA, Transfection, Cytotoxicity, Molecular biology, In vitro, Plasmid, Cell culture, Bioassay