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《JOURNAL OF LANZHOU MEDICAL COLLEGE》 1999-01
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Expression of a human TNF mutant in E.Coli and measurement of its bioactivity

WANG Xiao,SHEN Xingliang,YING Lianfang,et al.Military Medical Institute of Lanzhou Command,PLA,Lanzhou 730020  
Objective To synthesize a novel TNF and measure its bioactivity and expression. Method On the basis of analysis of TNF structure and the relationship between structure and function,a novel TNF coding sequence was synthesized by PCR technique and inserted into an expression plasmid.The novel TNF simultaneously made two alterations.Seven amino acids at amino terminus of TNF were deleted and ProSerAsp on 8910 sites were altered to ArgLysArg.Leu on 157 site was altered to Phe. Results By temperature induction,the transformed E.coli with the novel TNF expression plasmid produced high yield of novel TNF.Results showed that TNF protein content was 0.5 g/ml and the specific activity was 2.1105 BU/mg and the chemical and physical properties of novel TNF were different from those of nature TNF. Conclusions A novel TNF with bioactivity was constructed.
【CateGory Index】: R392-33
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