Study On Introducing Human Keratinocyte Growth Factor Into Carota And Rice

One of the hottest study in plant genetic engineering is that using the plant bioreactor to product the useful medicine or antibody. Compared with the microbe like E coli.,plant as a bioreactor is edible,cost low,needed no purification,and have the exact modification and process. Keratinocyte growth factor,which is composed with 194 amino acid and has three extrons and two introns,is one of the cell Mitogen-activated factors.KGF acts an important role in the growth and renew of epidermal cell.It can accelerate the bladder,nephridium,intestines and corneal epidermal wound healing.Also,it can alleviate the chemotherapy and irradiation induced injure.Some researches about the expression of KGF have been reported.Whereas it is few reported that to express KGF with plant bioreactor.So it is necessary to use the plant,such as rice and carota,to express the KGF.Firstly we synthesized the polynucleotide then obtained extron 1 of human KGF by the prolongation and PCR using the polynucleotide. Secondly using human lung DNA as template,we amplified extron 2 and extron 3 of KGF,then conjoined three extrons by overlap-extension PCR. Thirdly CAMV35s-KGF-NOS was obtained after ligation KGF with CaMV35S and NOS. Finally pBI1301-KGF was constructed after inserting CaMV35s-KGF-NOS into pBI1301.In this study,we introduced human KGF gene into rice and carota mediated by Agrobacterium.It is proved that KGF gene has inserted into the plant genome by the PCR and southern dot botting detection;it is also proved that the plant express the KGF by RT-PCR and western botting detection.Throught our study,we obtained the plant that express the KGF.It can be the groundwork for the application and exploitation of KGF transgene plant.