The Study Of Of Transgenes ScBADH And AFGF Enhances Stress-resistance Of Safflower Bioreactor

Recently, plant bioreactor has been investigated as a promising platform of producting medicinal proteins due to its intrinsic safety (it does not propagate mammalian viruses and pathogens), cost-effective bioprocessing that leads to lower production and downstream costs, and the capability for post-translation modifications. Plant bioreactor is a new research direction of biological pharmaceutical technology in recent years which has a broad market prospect to produce medicinal proteins.However,with the rapid increasing of world population and the global land salinization each year,how to solve the contradiction between food crisis and plant-bioreactor has become a pressing problem. Here,we show that the safflower transferred with both ScBADH (encoding betaineal dehydedehy drogenase from Suaeda corniculata) and aFGF (encoding acidic fibroblast growth factor from human) by Agrobacterium-mediated method.The transgenic safflower we get which can express aFGF protein and has greater resistance to the stress lay a firm material foundation for the creation of new resistance safflower bioreactor.And this also provide an important theoretical basis to alleviate the contradiction between the development of plant bio-reactor and the food crisis. The main results are as follow:1.Agrobacterium can infection safflower best when it was cultured in medium with acetosyringone100μM and spermidine1.μM. Add vitamin C50cysteine mg/L,L-Cysteine10mg/L,2g/L of carrageenan will reduce safflower explant Browning in tissue culture and prevention of vitrification, respectively.2.We first get the transgenic salfflower which transferred with ScBADH and aFGF by Agrobacterium-mediated method respectively. we achieve14and8transgenic plant which transferred with ScBADH gene and aFGF gene respectively by PCR analysis and the positive rate are3.15%å'Œ2.75%. The PCR results show that the target gene has transferred into salfflower DNA genome.3.Southern blot proved that ScBADH and aFGF have integrated into the salfflower genome. Integrated way seems to be asingle copy or multiple copies. Exogenous gene stably inherited in the generation.4.RT-PCR analysis showed that the transgenic safflower with wether ScBADH gene or aFGF gene can express on transcriptional level.5.SDS-page gel electrophoresis and Western blot hybridization analyses proved that aFGF expressed by transgenic safflower with aFGF gene has good immunogenicity.6. An examination of the stress-resistance characteristics demonstrated that the transgenic plants have higher badh activity and greater glycinebetaine accumulation due to the overexpressed of ScBADH gene which improved plant resistance to the stress.