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以墨西哥酸浆子叶作为外植体 ,与农杆菌EHA10 5 (含质粒 p130 1HBs)共培养 3天 ,通过延迟筛选的方法用潮霉素加压直接诱导成芽 ,抗性芽经诱导生根获得转化植株。经PCR、Southern点杂交等分子检测 ,证实目的基因已整合到墨西哥酸浆基因组中。ELISA检测证实了在墨西哥酸浆中表达的HBsAg具有较好的活性。初步比较了目的基因在转化植株、不同器官的表达水平差异。取注射初免 1次但抗体水平明显下降的Babl/c小鼠 ,口服饲喂加强免疫 ,有明显的特异性抗体回升反应。表明转基因墨西哥酸浆疫苗作为口服加强免疫具有重要的理论和实用价值。
The explants were cultured with Agrobacterium tumefaciens EHA10 5 (containing plasmid p130 1HBs) for three days and directly induced into shoots by hygromycin selection by delayed screening. The resistant shoots were induced to root and transformed Plant. By PCR, Southern blot and other molecular tests confirmed that the target gene has been integrated into the Mexican acid pulp genome. ELISA tests confirmed that HBsAg expressed in Mexican physalis had better activity. Preliminary comparison of the target gene in the transformed plants, the differences in the expression of different organs. Babl / c mice injected once with primary immunization but with a significant decrease in antibody levels were boosted orally with significant specific antibody response. This indicates that the transgenic Mexican physalis vaccine has important theoretical and practical value as an oral boost.