RESUMO
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.
Assuntos
Agrobacterium , Genética , Fator 1 de Crescimento de Fibroblastos , Genética , Vetores Genéticos , Genética , Plantas Geneticamente Modificadas , Genética , Metabolismo , Proteínas Recombinantes , Genética , Nicotiana , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To establish a high-frequency regeneration system of Astragalus and an aFGF transformation system.</p><p><b>METHOD</b>Cotyledon 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.</p><p><b>RESULT</b>All 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.</p><p><b>CONCLUSION</b>A high-frequency regeneration system of Astragalus and an aFGF transformation system were established.</p>
Assuntos
Humanos , Astrágalo , Genética , Metabolismo , Fator 1 de Crescimento de Fibroblastos , Genética , Metabolismo , Regulação da Expressão Gênica , Engenharia Genética , Métodos , Plantas Geneticamente Modificadas , Genética , Metabolismo , Transformação GenéticaRESUMO
AIM To explore the biotransformation of compound 7-(4-chlorbenzyl)-7,8,13,13a-tetrahydroberberine in the rabbit. METHODS Analyze the rabbit bile sample with HPLC, LC/MS and LC/NMR. RESULTS A metabolite and unchanged 7-(4-chlorbenzyl)-7,8,13,13a-tetrahydroberberine were found in the rabit bile, the metabolite was characterized and its structure was elucidated. CONCLUSION Compound 7-(4-chlorbenzyl)-7,8,13,13a-tetrahydroberberine is metabolized by demethylation at 10-OCH3 position.