بيان ژن انساني فعال كننده پلاسمينوژن بافتي (tPA) در گياه توتون تراريخت

Expression of human tissue Plasminogen Activator (tPA) in transgenic tobacco plants

In 1990s, applied molecular biology and plant biotechnology showed that plants can produce many drugs in high volume and very cheap. Plant system in safety, low cost, post-translation modifications and high volume production is better than classic systems (such as bacteria, yeast and animal cells). tPA (tissue Plasminogen Activator) drug protein especially and strongly bands to blood clots and preferably activates plasminogen into blood clots and then dissolve blood clots. tPA recombinant protein was used in stroke and cardiovascular diseases. In this report, the expression of tPA was evaluated in transgenic tobacco plants. This protein was expressed under the control of CaMV35S promoter and NOS terminator. A high expression sequence (kozak) and KDEL signal was linked to amino and carboxyl-terminal of tPA gene, respectively. The constructed cassette (pBI-tPA) transferred through Agrobacterium-mediated method to the genome of tobacco plants. The transgenic ones were selected on kanamycin (100 mg/L). Transgenic plants were evaluated by RT-PCR, SDS-PAGE, Zymography, Western blot techniques and all of them showed that tPA gene was transferred, actively expressed protein and recombinant tPA protein can dissolve clot bloods.