Transient expression of reteplase gene (rPA) under methyl jasmonat treatment in cucumber (Cucumis sativus L.)

Every year, millions of people are suffering from cardiovascular diseases. Thrombolytic and fibrinolytic therapies have shown results in the treatment of cardiovascular diseases and strokes. Tissue plasminogen activator (tPA) is a pivotal thrombolytic drug that plays a crucial role in cardiovascular disorders. The recombinant version of tPA, known as alteplase (rPA), has been designated as a significant breakthrough. Reteplase, a third-generation mutant of alteplase, has also emerged as a notable variant. The objective of the present study was to design a transient expression system to analyze the gene expression of rPA in cucumber plants. Moreover, the effect of elicitor methyl jasmonate (MeJA) was assessed on rPA gene expression. To do this, the rPA gene was amplified and subcloned into the ZYMV vector. The recombinant vector was then inoculated in cucumber leaves, and the transfected leaves were harvested at three, five, and ten days post-inoculation (DPI). An RT-PCR reaction was performed, and the desired band was observed in the inoculated lines. Furthermore, the quantity of rPA protein expression was assessed using ELISA. The results of the study demonstrated that the presence of methyl jasmonate led to an enhancement in the production of recombinant protein in the inoculated samples. The highest level of rPA gene expression was observed in the samples harvested on the tenth day post-inoculation with methyl jasmonate treatment which was ۴۵۰.۸۳ ng µg-۱ of total plant-extracted protein. For the first time, the rPA protein was successfully expressed under the influence of the methyl jasmonate elicitor in the cucumber plants.

Keywords:

Gene expression , Methyl jasmonate Elicitor , Pharming , Recombinant Protein , Reteplase , Transient expression