Pseudouridine sequencing using T. thermophilus tRNA mixture

The project focuses on understanding of the substrate specificity of T. thermophilus tRNA pseudouridine synthase D (TruD). In this project we generated a gene disruption strain in which the truD gene was disrupted using homologous recombination. We sequenced tRNA mixtures from both T. thermophilus wild-type and the truD gene disruptant to determine pseudouridine sites in tRNA. The NGS sequence revealed that TruD selectively modifies a subset of tRNA in vivo.