Optimizing of Gene Expression condition of Cyanovirin-LZN (anti-HIV protein) and Nanoformulation as Phytosome

Background: Antiretroviral (ARV) therapy were unable to control human immunodeficiency virus (HIV) infection completely, hence discovery of new vaccines as a novel formulation using effective microbicides (such as lectins) is essential. Recently, Cyanovirin-N (CVN) was considered as an important protein which is able to inhibit HIV infection. Methods and materials: Identified CVN gene was cloned into pET22b expression vector. The response surface methodology (RSM) was used to optimize the effect of expression parameters (temperature, IPTG concentration and time after induction). The gene expression levels (CVN and 16s rRNA: internal control) were calculated using real-time PCR. Following protein purification, it was formulated as phytosome complex (reflux method). Phytosome was evaluated using measurement of particle size distribution and zeta potential. Results: The new gene LZN-CVN (306bp) was identified and submitted in GenBank (KX673892). The maximum fold change level was predicted by RSM and according to the optimized expression condition (0.6mM IPTG, 29°C and 12h) recombinant protein (8KDa) was produced. The average particle size and zeta potential of phytosome complex were 132.5nm and -31.4mV, respectively. Conclusions: Recombinant protein was produced successfully using RSM. Based on results physical stability of LZN-phytosome complexe was confirmed.

Keywords:

Cyanovirin , Cloning , RSM , Phytosome , Human Immunodeficiency Virus