Construction of an expression vector for production of tissue plasminogen activator (t-PA) in a transgenic microalgae bioreactor

Microalgae systems have opened up new platforms for molecular farming in closed conditions. Recently, Dunaliella salina has been considered to be a suitable expression host for the production of recombinant proteins due to its significant advantages. It is a unicellular eukaryotic organism lacking cell wall and having post-translational processing for proteins which can grow in high salinity environments. To construct an expression vector, different fragments including Ubiquitin promoter, the gene of interest (t-PA; K2 and S domains), Omega leader sequences, KDEL, 6-Histidine tag, SYQ and Nos terminator (as the stop signal for gene transcription) were considered in upstream strategy. pCAMBIA 3301 and 1304 plasmids which contained bar gene (phosphinothricin resistance gene as a plant selectable marker) were used as the framework for construction of the expression vector and driving the expression of gene of interest. Further studies are undergoing to transform the constructed vector into D.salina.