Figure 5

Demonstration of co-transformation of heterologous genes. Paromomycin-resistant transformants were analyzed for the presence of the co-transformed, non-selectable DNA in the genome. PCRs were conducted using genomic DNA from transformants co-transformed with the pPsaD-GLuc (A), pHRLucP (B) or pHsp70A-GLuc (C) plasmids as template. The parental wild-type strain was analyzed as a control. Primers were specific for the heterologous gluc gene, and a 342-bp PCR fragment (Figure 3B-D) was expected in co-transformants. The rightmost lane shows a positive control using DNA of the co-transformed plasmid as the template. Lane M refers to the molecular weight marker. (D) Sequence obtained from the amplified and cloned gluc fragments. Primer positions, the stop codon (bold), and a BamHI restriction site (italics) are indicated.