Pennica D, Hayflick J S, Bringman T S, Palladino M A, Goeddel D V
Proc Natl Acad Sci U S A. 1985 Sep;82(18):6060-4. doi: 10.1073/pnas.82.18.6060.
A murine tumor necrosis factor (MuTNF) cDNA was isolated from a cDNA library prepared by using mRNA from the murine macrophage-like cell line PU5-1.8 induced with 4 beta-phorbol 12 beta-myristate 13 alpha-acetate. The cDNA encodes a polypeptide consisting of a 79 amino acid pre sequence followed by a mature MuTNF sequence of 156 amino acids. The 235 amino acid murine pre-TNF polypeptide is 79% homologous to the human pre-TNF protein. There is one potential N-linked glycosylation site on MuTNF, in contrast to human TNF, which lacks any such site. The MuTNF cDNA, when engineered for expression in Escherichia coli, was found to direct the synthesis of biologically active MuTNF as determined by its cytotoxicity against several transformed cell lines.
从小鼠巨噬细胞样细胞系PU5-1.8用4β-佛波醇12β-肉豆蔻酸酯13α-乙酸酯诱导后制备的cDNA文库中分离出小鼠肿瘤坏死因子（MuTNF）cDNA。该cDNA编码一种多肽，由79个氨基酸的前序列和随后的156个氨基酸的成熟MuTNF序列组成。235个氨基酸的小鼠前TNF多肽与人前TNF蛋白的同源性为79%。与缺乏任何此类位点的人TNF不同，MuTNF上有一个潜在的N-连接糖基化位点。当将MuTNF cDNA构建用于在大肠杆菌中表达时，通过其对几种转化细胞系的细胞毒性测定发现它能指导合成具有生物活性的MuTNF。
