الفهرس | Only 14 pages are availabe for public view |
Abstract Green biotechnology is the future of biopharmaceuticals production. The use of algae for the production of biopharmaceuticals is of particular interest in vaccine-based control programs. An attempt was made to express avian influenza virus (AIV) immunogens in algae because the virus is a serious economic and public health threat. A commercial eukaryotic expression system was modified to allow expression of AIV H5N1 hemagglutinin subunit 2 (HA2) in the microalga Chlamydomonas reinhardtii (C. reinhardtii). Codon-optimized AIV H5N1 HA2 sequences were synthesized and cloned (coHA2) into a cloning vector pUC-57. coHA2 sequences were then transferred to algal expression vector pChlamy_3/D- TOPO® then to C. reinhardtii strain cc-125. Proper nuclear integration was achieved in 16% of the screened transformants that survived selection in hygromycin-containing TAP media. coHA2 mRNA transcription was confirmed using RT-PCR, GFP expression using Confocal microscopy, AIV HA2 expression was confirmed using western blot analysis utilizing AIV H5N2 chicken antisera. These results warrant further investigation into immunogenic potential of the algae-expressed HA2 |