[ Objective] This study aimed to investigate the preparation of red component in secondary metabolites of recombinant Hansenula anomala strain N6076 and its GC-MS detection. [ Method] Thin-layer chromatography method ...[ Objective] This study aimed to investigate the preparation of red component in secondary metabolites of recombinant Hansenula anomala strain N6076 and its GC-MS detection. [ Method] Thin-layer chromatography method was applied for large-scale preparation of red component in the secondary metabolites of re- combinant H. anomala strain N6076. The red component was dissolved in anhydrous ethanol for GC-MS detection and chemical structure comparison in the data- base to identify its type. [ Resultl The red component is preliminarily identified as a quinane compound, while no compound with exactly the same structure as the red component has been found in WILEY, Nist and Nbs compound libraries. [ Conclusion] This study laid the foundation for further NMR structure identification of the obtained red component and investigation of the relationship between its structure and biolo#cal effects.展开更多
The sting of red imported fire ant （RIFA） could cause serious allergic response in fraction of people. These allergic reactions are mainly caused by its venom, especially venom allergen Sol i 1-4. To produce large a...The sting of red imported fire ant （RIFA） could cause serious allergic response in fraction of people. These allergic reactions are mainly caused by its venom, especially venom allergen Sol i 1-4. To produce large amount of RIFA venom allergen Sol i 4 for diagnosis of RIFA allergy and allergen-specific immunotherapy, the gene encoding this protein was amplified and cloned into the prokaryotic expression vector pET43, la. The recombinant plasmid was used to transform competent cells and the recombinant proteins were expressed in E. coll. SDS-PAGE and Western blotting analysis indicated that high-level expression of Sol i 4 protein was successfully achieved. Allergenic activity analysis of the recombinant allergen Sol i 4 was then performed on rabbit. The result showed that the recombinant protein obtained had significant allergenic activity. It indicated that the recombinant allergen Sol i 4 of RIFA venom was successfully expressed in E. coli, which provided foundation for further developing therapeutic and diagnosis reagents of RIFA allergy.展开更多
基金Supported by National Natural Science Foundation of China (30960006)Natural Science Fund of Xinjiang University (BS080120)Postdoctoral Station of Geography from Xinjiang University
文摘[ Objective] This study aimed to investigate the preparation of red component in secondary metabolites of recombinant Hansenula anomala strain N6076 and its GC-MS detection. [ Method] Thin-layer chromatography method was applied for large-scale preparation of red component in the secondary metabolites of re- combinant H. anomala strain N6076. The red component was dissolved in anhydrous ethanol for GC-MS detection and chemical structure comparison in the data- base to identify its type. [ Resultl The red component is preliminarily identified as a quinane compound, while no compound with exactly the same structure as the red component has been found in WILEY, Nist and Nbs compound libraries. [ Conclusion] This study laid the foundation for further NMR structure identification of the obtained red component and investigation of the relationship between its structure and biolo#cal effects.
基金supported by the Key Pro-gram Foundation of General Administration of QualitySupervision, Inspection and Quarantine of China(2005IK169).
文摘The sting of red imported fire ant （RIFA） could cause serious allergic response in fraction of people. These allergic reactions are mainly caused by its venom, especially venom allergen Sol i 1-4. To produce large amount of RIFA venom allergen Sol i 4 for diagnosis of RIFA allergy and allergen-specific immunotherapy, the gene encoding this protein was amplified and cloned into the prokaryotic expression vector pET43, la. The recombinant plasmid was used to transform competent cells and the recombinant proteins were expressed in E. coll. SDS-PAGE and Western blotting analysis indicated that high-level expression of Sol i 4 protein was successfully achieved. Allergenic activity analysis of the recombinant allergen Sol i 4 was then performed on rabbit. The result showed that the recombinant protein obtained had significant allergenic activity. It indicated that the recombinant allergen Sol i 4 of RIFA venom was successfully expressed in E. coli, which provided foundation for further developing therapeutic and diagnosis reagents of RIFA allergy.
