The Research On Acid Fibroblast Growth Factor Gene(a Fgf) Transfer Into Panax Ginseng By Agrobacterium-mediated

In this study, the plant prefer’s human a FGF gene was synthetic and the plant expression vector p CAMBIA1390-a FGF was constructed. The callus was induced by different culture media and culture conditions using 3-year-old ginseng root, stem, leaf as explants. The a FGF gene was transformed into Panax ginseng callus by Agrobacterium infection method. The resistance callus were acquired by hyg resistance selection. The transgenic ginseng callus was identified by PCR and RT-PCR method. The study laid the foundation for producing pharmaceutical proteins using ginseng as a bioreactor. The main results of this experiment are as follows:1. The plant expression vector construction experiments show: the plant preferred human a FGF gene was syntheticed and the plant expression vector p CAMBIA1390- a FGF was constructed successfully.2. Panax ginseng callus induction experiments showed that:(1) The induction rate was Panax ginseng stem > leaf > 3-year-old ginseng roots.(2) In the light of alternating dark conditions, ginseng callus induction rate was 96%, the color is green.(3) MS culture medium was more suitable for ginseng callus;(4) The optimum hormone of Panax ginseng callus induced was 2.4-D,secondly was NAA, BA, and KT.3. The genetic transformation experiments of Panax ginseng callus show: The infection condition of Panax ginseng callus was OD 0.6, infected 18 min in Agrobacterium, culture 60 h. and then received the most transformed callus tissue in selected medium.4. The screening and identification of the transgenic Panax ginseng callus results showed that: The transgenic Panax ginseng callus through Hyg screened has been identified by PCR and RT-PCR method in accordance with the optimum genetic transformation conditions. The a FGF gene has been transferred into Panax ginseng genome and transcription.5. The change of Panax ginseng saponin after a FGF gene transformation into Panax ginseng callus experiments show: in this study, the callus from transgenic and non-transgenic were choose for 3d, 7d, 14 d, 21 d to determine the accumulation of saponin content btween transgenic ginseng and non-transgenic callus.The results Xshow no effect on the content ginseng saponin after a FGF gene transfer into callus.