Expression of recombinant staphylokinase, a fibrin-specific plasminogen activator of bacterial origin, in potato (Solanum tuberosum L.) plants

One of the most dynamically developing sectors of green biotechnology is molecular farming using transgenic plants as natural bioreactors for the large scale production of recombinant proteins with biopharmaceutical and therapeutic values. Such properties are characteristic of certain proteins of bacterial origin, including staphylokinase. For many years, work has been carried out on the use of this protein in thrombolytic therapy. In this study, transgenic Solanum tuberosum plants expressing a CaMV::sak-mgpf-gusA gene fusion, were obtained. AGL1 A. tumefaciens strain was used in the process of transformation. The presence of the staphylokinase gene was confirmed by PCR in 22.5% of the investigated plants. The expression of the fusion transgene was detected using the β-glucuronidase activity assay in 32 putative transgenic plants. Furthermore, on the basis of the GUS histochemical reaction, the transgene expression pattern had a strong, constitutive character in seven of the transformants. The polyacrylamide gel electrophoresis of a protein extract from the SAK/PCR-positive plants, revealed the presence of a119 kDa protein that corresponds to that of the fusion protein SAK-mGFP-GUSA. Western blot analysis, using an antibody against staphylokinase, showed the presence of the staphylokinase domain in the 119 kDa protein in six analyzed transformants. However, the enzymatic test revealed amidolytic activity characteristic of staphylokinase in the protein extract of only one plant. This is the first report on a Solanum tuberosum plant producing a recombinant staphylokinase protein, a plasminogen activator of bacterial origin

Nuclear DNA endoreplication and plastid index in mesophyll of some dicotyledonous species

Cytophotometric studies of nuclear DNA content after Feulgen procedure indicate that in mesophyll of all the seven studied species the highest nuclear DNA endoreplication level occurs in II or III leaf and it varies for particular species. No differences were found in nuclear DNA endoreplication dynamics between the basal and apical parts of the leaf blade. Chloroplast number per cell generally decreases in the successive leaves, and the plastid index is the smallest in the first (oldest) leaves, being similar in both zones. In four species chloroplast number and plastid index show relatively low negative correlation with nuclear DNA contents (expressed as endoreplication index), in two species this correlation is positive, and one species displays very low r value