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Gene sequence of porcine tumor necrosis factor alpha.

We report genomic DNA sequence for porcine tumor necrosis factor alpha. The gene segment was obtained by PCR (1) from porcine genomic DNA using the primers: 5' -GCGGATCCATGAGCACTGAAAGCATGATC -3 ' (sense) and 5'-GCCGAATTCAGGGCAATGATCCCAAAGT-3' (antisense). Amplification was performed in a homemade thermal cycling apparatus (2). Primers were based on published human sequence (3) from the extremes of known coding regions. Restriction sites were included at the 5' ends as indicated (underlined). A PCR product of ~ 1.8 kb was excised and purified from a .7% agarose gel, cleaved with EcoRI and BamHI, cloned into pUC19, and further subcloned into M13 vectors for conventional dideoxy sequencing in both strands. The resulting sequence (BamHI-EcoRI fragment, 1806 bp) is shown in Figure 1. Coding regions of this sequence differ from the PCR-derived partial cDNA sequence of Pauli and co-workers (4) at three positions: C not T at 1657, A not G at 1768, and T not C at 1771. Regions corresponding to 136-156 and 1766—1802 of this sequence were primer-derived in the sequence of Pauli and co-workers. Curiously, the 3' nucleotide (G) of their sense primer for cDNA amplification differed from their reported cDNA sequence (an A corresponding to our position 156). None of these differences alter the predicted translation, however.