[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF wa...[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed （ Brassica campestris L. ） by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows： explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.展开更多
Among 27 filamentous fungi, a strain Aspergillus niger An76 gave the highest siderophore yield even when cultured on natural medium or minimal medium containing 1 mmol L^-1 Fe^3＋. Whereas for most other strains, the ...Among 27 filamentous fungi, a strain Aspergillus niger An76 gave the highest siderophore yield even when cultured on natural medium or minimal medium containing 1 mmol L^-1 Fe^3＋. Whereas for most other strains, the capacities of siderophore production was repressed as the concentrations of Fe^3＋ was higher than 20 ~tmol L^-1. Lower ferric iron concentrations were more conducive to siderophore synthesis for those strains. The An76 siderophore was partially purified through a series of chromatography steps using Sephadex LH20 and CM-Sepharose Fast Flow columns. Its structural characteristics differed from typical siderophores, such as catecholates or hydroxamates, and no amino acids or peptide bonds were detected. It may therefore represent a new type of siderophore structure. TBA assay and ESR trapping studies showed that An76 siderophore had remarkable scavenging activity of hydroxyl free radicals in vitro. The antibiotic activities of siderophores produced by An76 were tested against 29 bacterial strains, including plant, animal, and human pathogens. The growth of 14 bacterial strains could be completely inhibited at a siderophore concentration of 8 mg ml^-1 with an incubation time of 30 min. Evidence suggests that the An76 siderophore has potential value as an antioxidant and a biocontrol agent against pathogenic microorganisms.展开更多
基金Supported by Bioreactor Important Special Item of 863-Program inthe "Eleventh Five-Year" Plan (No. 2007AA100503)Science and Technology Development Key Plan of Jilin Province( No.20070922)+1 种基金Cultivation Fund of Scientific and Technical Innovation Project Major Program of Higher Education Institutions ( No.70S018)Science and Technology Plan of Changchun City (No.06GG150)~~
文摘[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed （ Brassica campestris L. ） by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows： explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.
基金This work was supported by the National Natural Science Foundation of China （No. 30470051）, the Major State Basic Research Development Program of China （973, No. 2004CB7197）, and the Natural Science Foundation of Shandong Province （No. Y2004D07）.
文摘Among 27 filamentous fungi, a strain Aspergillus niger An76 gave the highest siderophore yield even when cultured on natural medium or minimal medium containing 1 mmol L^-1 Fe^3＋. Whereas for most other strains, the capacities of siderophore production was repressed as the concentrations of Fe^3＋ was higher than 20 ~tmol L^-1. Lower ferric iron concentrations were more conducive to siderophore synthesis for those strains. The An76 siderophore was partially purified through a series of chromatography steps using Sephadex LH20 and CM-Sepharose Fast Flow columns. Its structural characteristics differed from typical siderophores, such as catecholates or hydroxamates, and no amino acids or peptide bonds were detected. It may therefore represent a new type of siderophore structure. TBA assay and ESR trapping studies showed that An76 siderophore had remarkable scavenging activity of hydroxyl free radicals in vitro. The antibiotic activities of siderophores produced by An76 were tested against 29 bacterial strains, including plant, animal, and human pathogens. The growth of 14 bacterial strains could be completely inhibited at a siderophore concentration of 8 mg ml^-1 with an incubation time of 30 min. Evidence suggests that the An76 siderophore has potential value as an antioxidant and a biocontrol agent against pathogenic microorganisms.
