On the TATA box and transcriptional start site of gE gene of pseudorabies virus: a comparison of three methods

Using the upstream region of glycoprotein E gene of pseudorabies virus (PRV) as a model, a method based on reverse transcriptase-polymerase chain reaction (RT-PCR) for the determination of transcriptional start site was developed. The conventional primer extension method was used to determine the start site. Comparing the results obtained by these two methods, a good agreement on the location of start site was achieved. In addition, a computer program was used to predict the transcriptional start site, and the predicted site was found to be close to the two sites obtained by experiments. Based on the transcriptional start site data and on the general knowledge of the eukaryotic gene, the TATA box of gE gene was assigned at ∼33 bp upstream to the start site.