Construction Of Plant Expression Vector Of HEGF And Genetic Transformation Into Plants By Agribacterium Tumefaciens

The human epidermal growth factor (hEGF) is a powerful mitogen for a wide variety of cells in culture and in vivo, which is a kind of tyrosine kinase leading to itself phosphorylation and cells proliferation. It is often applied to treat burns, scald, cornea transplant, and intensively promotes the proliferation and growth at very low dosage in many types of cells especially in protecting and repairing the alimentary canal epithelium cells and wound healing. Furthermore, the human EGF can be also widely used in the cosmetic. It has big latency economic value because it has special and potential biological effects. Plants have many advantages compared with traditional systems for the production of biopharmaceutical proteins. These include the low cost of production, rapid scalability, the absence of human pathogens and the ability to fold and assemble complex proteins accurately.It becomes one of the research hotspots.In the present study, The hEGF gene sequence has been designed and chemically synthesized according to enregistered hEGF sequence and the codons bias in plant. Amino acid GPGP and SEKDEL as endoplasmic reticulum orientation sequence were added in 3' end, in order to improve protein expression. Expression vectors pSH-hEGFS, pFCor15FGN, pCambia-Pnos-PNN-hEGFS and pFCor15FGN-hEGF suitable for expression in plant has been constructed. The CaMV35S promoter and NOS terminator fragments were syncretized with reporter GUS gene and targeted gene. The selectable maker gene was neomycin phosphotrasferase gene, which allowed selection with kanamycin. Both tobacco and petunia were been transformed by Agrobacterium tumefaciens . The buds were selected by kanamycin. The results of GUS histochemical tinction and hEGF gene PCR indicated that hEGF gene have been integrased into genomes of tobacco and petunia respectively help to develop the bioactor producing hEGF using transgenic plants.