百脉根高频再生体系建立和猪瘟病毒E2基因的遗传转化

摘要

在建立百脉根（Lotus corniculatus L .)高频再生体系的基础上，通过农杆菌介导，将猪瘟病毒（CSFV)表面抗原E2基因转化进入百脉根，获得转基因阳性株系5个。并对遗传转化体系中关键性因素进行优化，结果表明，最优化体系为子叶预培养3 d后，用OD600nm为0.5±0.05的菌液侵染30 min，共培养3 d并添加AS（乙酰丁香酮)，选择培养基中卡那霉素浓度为50 mg/L，可获得理想的转化效率。为抗猪瘟可食性疫苗的研制奠定基础。%Based on the established regeneration system of Lotus corniculatus L. with high frequency and mediated by Agrobacterium tumefaciens,5 lines of transgenic Lotus corniculatus L. with E2 gene of classical swine fever virus(CSFV) were obtained. Several key factors affecting genetic transferring system were optimized during E2 genetic transformation. The results showed that the optimal system was pre-cultivated cotyledon for 3 days and infected with Agrobacterium at the concentration of OD600nm=0.5±0.05 about 30 min. After co-cultured for 3 days, 100μmol/L AS was added in culture medium containing 50 mg/L Kanamycin. Under this system, ideal efficiency of genetic transformation was achieved. It will lay a foundation for edible vaccine of resisting swine fever.