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生菜遗传转化受体系统的建立及鲑鱼降钙素基因的导入 被引量:10

Establishment of Genetic Transformation System of Lettuce and Introducing of Salmon Calcitonin Gene
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摘要 以散叶生菜大速生(Lactucasativavar.capatataL.)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+0.1mg/L6 BA+0.05mg/LNAA;抗生素敏感性试验表明,筛选培养基中适宜的卡那霉素选择压为75mg/L,抑菌剂羧苄青霉素的适宜质量浓度为300mg/L;通过根癌农杆菌介导的叶盘法将鲑鱼降钙素(sCT)基因转入大速生散叶生菜,PCR检测转化率达25.4%. Studies on establishment of receptor system of gene transformation of lettuce (Lactuca sativa var. capatata L.) are reported. The effects of different hormone concentration on shoot regeneration from cotyledon explants are investigated. The results show that MS medium supplied with 0.1 mg/L 6-BA and 0.05 mg/L NAA is the suitable shoot regeneration medium. The experiment on sensitivity of cotyledon to kanamycin and carbenicillin shows the suitable kanamycin concentration for selecting transgenic tissue is 75 mg/L and the carbenicillin concentration is 300 mg/L. Salmon calcitonin (sCT) gene is transferred into lettuce mediated by Ti plasmid of Agrobacterium tumafaciens. 118 lettuce whose resistance kanamycin is obtained after sifted on shoot regeneration medium and root induce medium supplied with 75 mg/L Km; PCR amplification of genomic DNA from kanamycin resistance lettuce leaf tissue shows sCT gene has been integrated into the genome of transgenic lettuce.
出处 《烟台大学学报(自然科学与工程版)》 CAS 2004年第2期116-121,共6页 Journal of Yantai University(Natural Science and Engineering Edition)
基金 北京市重点实验室资助项目(951890200).
关键词 遗传转化 鲑鱼降钙素基因 生菜 异源蛋白 激素 gene transformation salmon calcitonin gene lettuce
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