In order to investigate the inhibitory effect of microwave exposure on the import of mitochondrial transcription factor A (Tfam), we successfully created a recombined expression vector Tfam-pTNT, which was then placed into rabbit coupled reticulocyte lysate system to be translated into the precusor of Tfam (pre-Tfam) labeled with S,35, methionine. After exposing the primary cultured cortex neurons to microwave for 15min, we isolated the mitochondria from these treated neurons at 0 h, 6 h, 12 h, 24 h and 48 h after microwave irradiation. Through import assay, it was found that microwave exposure obviously decreased the import of the pre-Tfam from cytoplasm into mitochondria in a time-depend manner. The results indicate that microwave exposure can inhibit the import of Tfam, which reveals a novel mechanism that microwave irradiation disturbs the energy metabolisms in neurons.