口蹄疫病毒P12A-3C免疫原基因在百脉根中的遗传转化与表达

Transformation and expression of P12A-3C gene of foot and mouth disease virus(FMDV) into transgenic plant Birdsfoot trefoil

目的利用植物反应器研究口蹄疫转基因植物疫苗是近些年来科学家研究的热点,本研究以豆科牧草百脉根为转化受体,将口蹄疫病毒P12A-3C基因通过根癌农杆菌介导法导入百脉根基因组。方法百脉根外植体经过浸菌,抗性培养基上愈伤、出芽和生根等阶段。结果最终获得了转基因抗性植株。结论对转基因植株进行PCR、RT-PCR检测,表明外源基因整合在植物染色体基因组,并且具有转录活性,ELISA检测表明,转基因植株表达出外源目的蛋白。

In the present study, the P12A-3C gene of foot and mouth disease virus （FMDV） was transformed to the transgenic plant Birdsfoot trefoil mediated with Agrobacterium tumefaciens GV3101. A aeries of experiments including the callus culture, shooting and rooting regeneration on the selective medium were performed and the transgenic plants were generated on medium containing 50 mg/L Kan. The presence of the immunogen gene was tested by RT-PCR. It was demonstrated that an approximate 3 kb DNA fragment was amplified in the transgenic plant host ceils. However, the DNA fragment of P12A-3C gene was absent in the non-transformed transgenic plants. As demonstrated by RT-PCR analysis, the transcriptional activity could be detected in the transgenic plants, but it was absent in the non-transformed plants. It was also found that the exoge- nous target protein could be detected in the transformed plants as demonstrated by ELISA assay.