Transient expression of CCL21 chemokine in chickpea via agroinfiltration

IntroductionChemokine (C-C motif) ligand 21 (CCL21) has anti-tumor efficacy and used for immunotherapy based on cytokinins against cancer cells. Immune potentiating of CCL21 chemokine via DC and stromal cell-based approaches for effective recruitment and activation of APC and T cells for promotion of antitumor activity in lung cancer was evaluated. Plants are now gaining widespread acceptance as a general platform and as bioreactors for the large scale production of recombinant proteins. However, the technique suffers from major drawbacks such as low expression level and long time required for the production of the recombinant protein in plant tissues. The goal of the present study was to investigate the possibility of expressing the CCL21 protein in Cicer arietinum via agroinfiltration. In agroinfiltration, the suspension of Agrobacterium tumefaciens harboring the gene(s) of interest is infiltrated into plant leaves using a needle-free syringe. This technique has been carried out in a variety of plants with different experimental purposes and it was simple, cost effective and rapid procedure. Chickpea contain several minerals and phytochemicals components for human health, for example it has selenium, that plays essential roles in liver enzyme function, and helps detoxify some cancer-causing compounds in the body, Also selenium prevents inflammation and also decreases tumor growth rates. So cheakpea known as a valuable plant for the production of recombinant proteins chickpeas also contains folate, which plays a role in DNA synthesis and repair, thus preventing the formation of cancer cells from mutations in the DNA. Also it has saponins, which are phytochemicals that prevent cancer cells from multiplying and spreading throughout the body. High-fiber intakes from fruits and vegetables like chickpeas are associated with a lowered risk of colorectal cancer. Therefore, in this study, ccl21 gene of human was synthesized and transiently expressed in Cicer arietinum. Materials & MethodsTo examine the possibility of expressing the CCL21 protein in Cicer arietinum, a cDNA fragment encoding the ccl21 gene was synthesized de novo, modified with a Kozak sequence, a C-terminal hexa-histidine (6His) tag, and an endoplasmic retention signal (SEKDEL). The construct was subcloned into vector pBI121 and Agrobacterium colonies were verificated by PCR test. The resulting ccl21 plasmid was agro-infiltrated into Cicer arietinum. The relative gene expression of recombinant pla