Tg(Nefl-TNF*)K3Gkl

The 7-kb murine Nefl gene, containing 1.7 kb of 5'-flanking sequences that confer neuronal specificity was fused to human TNF genomic sequences containing a deletion mutation of the codons encoding the 1st to the 12th amino acids of the mature 17-kDa TNF protein. This mutant TNF gene has previously been shown to produce a transmembrane TNF protein that remains bioactive against cellular targets in vitro and in vivo. The genomic DNA was then ligated to the 0.77-kb EcoRI-Sal1 fragment containing the 3' UTR and polyadenylation site of the beta-globin gene, which replaces TNF 3' UTR sequences, thought to negatively regulate mRNA stability and translational efficiency. In this transgenic line, exogenous TNF protein production could be demonstrated by positive imnmunostaining of neurons in the spinal cord and brain sections using a polyclonal anti-human TNF Ab. Transgenic lines TgK1, TgK11 and TgK14 were also created. (J:128072)