Construction of the expression plasmid pHu (AnTH1). The figure illustrates the process started from Poly-A mRNA purification from cell lines, followed by the RT-PCR of respective cDNA fragments. The amplified cDNA TH1 antagonist (AnTh1) contains in its extremes the sequences for NcoI and BamHI restriction enzymes suited for cloning in the pPTV-1 vector (left at the bottom of diagram). In the center is indicated the nucleotide sequence (ccatatgatg) coding for the linking peptide. The final genetic construction is shown at the bottom right.