口蹄疫抗原决定簇融合基因转化生菜的研究

Transformation of Lettuce with FMDV Epitopes Fused Gene Mediated by Agrobacterium

植物疫苗具有生产简单、成本较低、使用安全方便、易贮存等优点,是目前植物基因工程的一个研究热点。本实验通过农杆菌介导的遗传转化,以O型和A型口蹄疫抗原决定簇融合基因O21-O14-A21-HBcAg转化生菜。通过筛选,获得潮霉素抗性愈伤组织,经胚状体再生得到56棵抗性苗。对部分抗性植株进行PCR和PCR-Southern杂交检测,证实目的基因已经成功整合到生菜基因组中。RT-PCR检测初步表明,O21-O14-A21-HBcAg基因可以在生菜中正常转录表达。

Plant-derived vaccine is becoming one focus of plant gene engineering, which has advantages such as easy production, low cost, convenient and safe use. In this experiment, the fused gene O21-O14-A21 containing both type O and A FMDV epitopes was introduced into lettuce mediated by agrobacterium. The hygromycin-resistant lettuce calli were obtained by screening on the medium containing antibiotic. Fifty six hygromycin-resistant lettuce seedlings were regenerated. The results of PCR and PCR-Southern blotting indicated that the fusion gene were successfully integrated into the genomes some of transformed lettuce plants. Expression of O21-O14-A21 gene in transformed lettuce plants was proved by RT-PCR analyses.