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生菜遗传转化体系的建立及口蹄疫抗原决定簇融合基因的导入
Establishment of Genetic Transformation System of Lettuce and Introducing of FOMD Antigenic Determinant Fused Gene
【摘要】 以散叶生菜大速生为试材,以MS为基本培养基,采用不同激素配比,确定生菜高效诱芽培养基为MS+1.5 mg/L 6-BA+0.2mg/L IAA;抗生素敏感性试验表明,筛选培养基中适宜的潮霉素选择压为20 mg/L,抑菌剂羧苄青霉素的适宜质量浓度为300 mg/L;通过根癌农杆菌介导的叶盘法将携带O型和A型口蹄疫抗原决定簇融合基因O21-O14-A21-HBcAg转入大速生散叶生菜,PCR检测转化率达50%。
【Abstract】 Studies on establishment of receptor system of gene transformation of lettuce(L act uca sativa var.capatata L.) are reported.The effects of different hormone concent ration on shoot regeneration from cotyledon explants are investigated.The results show that MS medium supplied with 1.5 mg/ L 6-BA and 0.2 mg/ L IAA is the suitable shoot regeneration medium.The experiment on sensitivity of cotyledon to hygromycin B and carbenicillin shows the suitable hygromycin concentration for selecting transgenic tissue is 20 mg/ L and the carbenicillin concentration is 300 mg/ L.Fused gene O21-O14-A21 containing both type O and A FMDV is transferred into lettuce mediated by Ti plasmid of Agrobacterium tumafaciens.56 lettuce whose resistance hygromycin is obtained after sifted on shoot regeneration medium and root induce medium supplied with 20 mg/L Hyg;PCR amplification of genomic DNA from hygromycin resistance lettuce leaf tissue shows O21-O14-A21 gene has been integrated into the genome of transgenic lettuce.
【Key words】 FMDV antigenic diterminant; agrobacterium tumefaciens; lactuta sativa; genetic transformation; gene expression;
- 【文献出处】 平原大学学报 ,Journal of Pingyuan University , 编辑部邮箱 ,2006年02期
- 【分类号】Q943.2
- 【被引频次】3
- 【下载频次】90