Expression Analysis Recombinate Human Epidermal Growth Factor Gene In Transgenic Plants

Human epidermal growth factor(hEGF) is a small peptide hormone which stimulates the proliferation or differentiation of a wide variety of cell.Its commercial applications rangs from wound healing and inhibition of gastric acid secretion to wool harwesting.Molecular farming is the production of pharmaceutically important and commercially valuable proteins in plants.Molecular farming in plants has the potential to provide virtually unlimited quantities of recombinant proteins for use as diagnostic and therapeutic tool in health care and the life sciences.This demonstrates the promise of using transgenic plants as bioreactors for the molecular farming of recombinant therapeutic,including vaccines,diagnostics,such as recombinant antibodies,plasma proteins,cytokines and growth factors.In this study,Plant expression vector pSH-hEGFS was analysed by PCR.Transient expression of pSH-hEGFS was analysed in lettuce via Agroinfiltration-the infiltration.The vector containing hEGF gene was transformed into tobacco by Agrobacterium tumefaciences. The expression of hEGF was studied.The results are as the follows:1.Transient expression of pSH-hEGFS was analysed in lettuce via Agroinfiltration-the infiltration of Agrobacterium tumefaciens into intact plant leaves.GUS activity reached its maximum (29.2U/g) within 4-6 days following agroinfiltration and decreased slowly.2.Regenerated plantlets were obtained via co-culture,shoot inducing and root inducing. Transgenic plants were conformed by the GUS histochemistry analysis,PCR analysis and RT-PCR analysis.The results indicated that the hEGF gene had been integrated into the genome of tobacco and expressed.3.The hEGF content in transgenic tobacco leaves reached to levels of 0.003%of the total soluble protein in leave by ELISA assay.4.The botanical character of transgenic plants,such as plant type,leaf shape,leaf angle related to stem,stalk circumference,internode length,inflore scence,flower color,showed almost as same as these character of control plants.It indicated that expression of the hEGF gene did not effect the growth of tobacco.