TLMA2993 peptide (N'-TLMALELKGKLLLAGLAPSAFLPLSFPEGL-C') which was designed by a computer program (MIMETIC) inhibited the activity of HIV-1 reverse transcriptase in a cell-free system. Therefore, we constructed a TLMA2993 expression vector containing an artificial cDNA for TLMA2993 to generate the peptide in cells. The cell lysate of transfected U937 cells contained a detectable level of TLMA2993 peptide using competitive ELISA. The transfectants were resistant to HIV-1 infection due to expression of TLMA2993 peptide in the cells. The use of MIMETIC to design an inhibitory peptide to any intracellular target molecule, followed by transfection of the artificial cDNA for the peptide, could afford a new approach for treatment and/or prevention of viral infection.