OBJECTIVETo establish a high-frequency regeneration system of Astragalus and an aFGF transformation system.

METHODCotyledon node of the Astragalus explants was used for organogenesis to establish a high-frequency regeneration system. GV3101 was used to transform cotyledon node, and aFGF gene was introduced into Astragalus, renewable strain was detected by PCR.

RESULTAll cotyledon node was explants, adventitious buds were induced in the medium of MS +2.0 mg x L(-1) BA +0.5 mg x L(-1) IBA, the root was taken in the medium of 1/2MS +5.0 mg x L(-1) NAA to give a high frequency regeneration system. All cotyledon node was precultured in medium for 3 days and infected with Agrobacterium (A600 0.3) for 10 min and then cocultured for 2 days. The aFGF gene was confirmed to transform into genome of Astragalus.

CONCLUSIONA high-frequency regeneration system of Astragalus and an aFGF transformation system were established.

Astragalus Plant ; genetics ; metabolism ; Fibroblast Growth Factor 1 ; genetics ; metabolism ; Gene Expression Regulation ; Genetic Engineering ; methods ; Humans ; Plants, Genetically Modified ; genetics ; metabolism ; Transformation, Genetic

Acid fibroblast growth factor (aFGF) has great potential in clinical application, but it is very expensive. In order to reduce the cost of production and to make full use of the merits integrated with plant bioreator, we have explored the aFGF in transgenic Tobacco expression. AFGF gene was inserted into plant expression vector pBI121; the acquired plants contained aFGF gene expression vector pBI121-TOAB-aF. Using Agrobacterium-mediated gene transformation of Tobacco and using transgenic Tobacco containing kanamycin and cephalosporin culture medium, we obtained kanamycin resistant transgenic Tobacco plants. PCR detection, RT-PCR detection and Western blot detection confirmed that foreign genes were successfully expressed in Tobacco. These data could serve as a theoretical foundation on which to use the plant bioreactor for production of aFGF.
Agrobacterium ; genetics ; Fibroblast Growth Factor 1 ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Tobacco ; genetics ; metabolism