牛皮蝇HA抗原基因转化紫花苜蓿的研究

以“甘农1号”紫花苜蓿(Medicago sativa)7 d苗龄子叶和下胚轴为受体材料，建立了高效的苜蓿再生体系和遗传转化体系，筛选出MS+2,4 D 2.0 mg/L+6 BA 0.5 mg/L和MS+6 BA 0.5 mg/L+NAA 0.03 mg/L+GA3 2.0 mg/L为苜蓿子叶愈伤组织诱导和分化的适宜培养基；探讨了农杆菌浓度OD600约0.5、感染时间8~10 min、共培养时间2 d为适宜的转化条件。采用农杆菌介导法将Hyperdomin A（HA）基因导入紫花苜蓿，经PCR检测和Southern分子杂交分析，结果表明HA基因已经整合到苜蓿基因组中，可为牛皮蝇蛆病可食性疫苗的研究提供理论基础。

A study on transformation of Hypodermin A gene into Medicago sativa

The high efficiency regeneration and genetic transformation system of alfalfa （Medicago sativa）was established by using the cotyledons as explants from the sterile 7 days old seedlings of M.sativa cv. Gannong No.1. The optimal medium for callus induction and regeneration from Gannong No.1 cotyledons was MS+2,4 D 2.0 mg/L+6 BA 0.5 mg/L and MS+6 BA 0.5 mg/L+NAA 0.03 mg/L+GA3 2.0 mg/L respectively. In this study, the efficiency transformation system included agrobacterium concentration OD600≈0.5, infection time 8-10 min and co culture time 2 d. The transgenic plants were identified by PCR and Southern blot techniques. The results indicated that the Hyperdomin A （HA） gene had been transformed into the genome of Gannong No.1 alfalfa.