Expression of The Human Acidic Fibroblast Growth Factor in Transgenic Tomato and Safety Assessment of Transgenic Lines

摘要

The establishment of transgenic tomato (Solanum lycopersicum) lines expressing human acidic fibroblast growth factor (haFGF) was reported. A nonantibiotic selection involving the manA gene from E. coli that confers resistance to mannose was applied. The transgenic nature of four selected tomato lines cultivar Bela was proved with PCR analysis. The expression of the gene for haFGF was confirmed with RT-PCR analysis and the protein synthesis was quantified with ELISA. A step of purification of haFGF from plant tissue was also performed and the probes were subjected to immunodetection with anti-haFGF and anti-His-tag antibodies. The portion of the recombinant protein out of the total soluble protein extracted from leaves was 0.0015%. In order to evaluate the risk of unintended negative effects of the ripe transgenic tomato fruits, toxicity safety assessment with laboratory mice was carried out including weight analysis, morphophysiological indices and blood parameters measurements. Our data showed that the transgenic fruits as a novel modified product do not pose a risk of intoxication upon the treated animals. Allergenicity evaluation of both recombinant proteins haFGF and phosphomannose isomerase, the product of the selectable marker gene, via alignment of their sequences with those of known allergens did not reveal any sufficient similarity.