Figure 3

A Schematic representation UL regions of the DEV genome containing the UL31 gene and strategy for construction of the expression plasmid pET 32-UL31. B PCR product of the fragment of DEV UL31 detected by 1% agarose gel electrophoresis. Lane M, DNA marker; Lane 1, PCR product of the DEV UL31. C DEV UL31 gene encoding DNA sequence was cloned into pET 32a(+) prolaryotic expression vector as described in materials and methods. The construct was digested with two restriction enzymes. M, DNA marker; Lane 1, Bam HI generating one restriction fragment; Lane 2, Bam HI and Hind III generating two restriction fragments. D Induction of the His-tagged UL31 fusion protien in E. coli. Plasmid pET-UL31 was transformed into bacteria. Bacteria were grown in the absence (lane 1) or the presence (lane 2) of IPTG. The fusion protein was purified as described in Methods (lane 3). Molecular mass markers (in kDa) are shown to the right (lane M). The arrowhead indicates the induced UL31 fusion protein.