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Studies on Construction of Plant Expression Vector of Recombinant LBM2eHBc+ Gene and Tomato Transformation

Abstract

禽流感病毒一直是家禽和家畜健康养殖的巨大威胁,甚至威胁着人类的健康,人们也一直在研究各种形式的疫苗来应对禽流感病毒的威胁.番茄作为生物反应器应用于动物疫苗生产具有独特的优点,本研究优化了番茄子叶的再生体系,同时构建了lbM2EHbC+融合基因的植物表达载体并对番茄进行了遗传转化.结果表明在玉米素(zT)浓度为0.5 Mg/l时番茄子叶外植体的芽再生率达到93.33%;测序结果证明植物表达载体PbI121-lbM2EHbC+构建成功;利用农杆菌介导的转化方法获得再生抗性苗34株,经PCr检测,阳性率为75.0%,本研究结果为进一步开发禽流感口服疫苗奠定了基础.Avian influenza virus(AIV) infect almost all wild bird and domestic poultries and then decimate poultries,even the threat of AIV overhangs human.It also has been researching a variety of vaccines to combat the AIV threat.Tomato as a bioreactor used in animal vaccine production has a unique advantage.This study optimized the regeneration system of tomato cotyledon,while construct LBM2eHBc + gene plant expression vector,and then carried out the genetic transformation of tomato.The results show that the bud regeneration rates of tomato reach the highest 93.33% when MS medium supplemented 0.5 mg/L Zt+ 1 mg/L IAA;sequence results show that plant expression vector pBI121-LBM2eHBc + constructed successfully.A total of 34 putative transgenic tomato plants were obtained by using of Agrobacterium-mediated transformation method,and the positive rate was 75.0% by PCR detection.The results laid a foundation for the further development of oral AIV vaccines.福建省科技厅重大项目子课题(2006NZ0003-2);福建省教育厅A类科技项目(JA09168

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