条斑紫菜叶状体细胞对3种抗性筛选剂敏感性的研究  

The Susceptibility of Somatic Cells from Thalli of Laver Porphyra yezoensis to Three Selective Reagents

作　　者：王萍[1] 金俊[1] 滕亚娟[1] 李信书[1] 陈平[1] 周俊 

机构地区：[1]淮海工学院海洋学院江苏省海洋生物技术重点实验室,江苏连云港222005 [2]句容市同力生物科技有限公司,江苏句容212402

出　　处：《水产科学》2013年第12期691-695,共5页Fisheries Science

基　　金：江苏省海洋生物技术重点实验室开放课题(2005HS011);淮海工学院引进人才科研启动基金资助项目(KK01073)

摘　　要：为探讨条斑紫菜叶状体细胞对3种抗生素的敏感性，以确定条斑紫菜遗传转化后代适宜的抗性选择质量浓度，将叶状体用海螺酶解离获得原生质体，在培养皿中20℃培养3 d后，在培养液中分别加入G418（0、10、50、100、200、300 mg/L）、卡那霉素（0、50、100、300、500、800 mg/L）或氯霉素（0、5、15、35、65、100 mg/L ），观察细胞分裂与死亡率、小紫菜比率，进行显著性检验。试验结果表明，条斑紫菜叶状体细胞对3种抗生素的敏感性存在显著差异，对抗生素的敏感性为氯霉素＞卡那霉素＞ G418。在条斑紫菜叶状体遗传转化用G418、卡那霉素或氯霉素作抗性筛选时，筛选质量浓度分别为300、50 mg/L及15～35 mg/L。The susceptibility of somatic cells of thalli of laver Porphyra yezoensis to three antibiotics was studied in order to screen reliable selection concentrations for recovery of transformation offsprings of the laver .The protoplasts isolated from thalli by seashell enzymes were cultured in medium at 20 ℃ for 3 days , and then transferred to medium with G418（0 ,10 ,50 ,100 ,200 ,and 300 g/L） ,kanamycin（0 ,50 ,100 , 300 ,500 ,and 800 mg/L） or chloramphenicol （0 ,5 ,15 ,35 ,65 ,and 100 mg/L） .The rate of cell division , mortality ,and rate of thallus formation were observed with significant difference test .The results showed that there were significant differences in susceptibility of the somatic cells to three antibiotics at various concentrations .It was found that the susceptibility of the cells from the thalli to the antibiotics was arranged as chloramphenicol 〉 kanamycin 〉 G418 .As selective reagents for recovery of transformation offspring of the thallus ,the concentration was selected a 300 mg/L for G418 ,50 mg/L for kanamycin and 15~35 mg/L for chloramphenicol .

关 键 词：条斑紫菜 叶状体 抗生素 筛选标记 

分 类 号：S968.431[农业科学—水产养殖]