| 摘要: |
| 对外源基因导入大菱鲆(Scophthat musmaximus)的两种方法进行了研究,采用经过改进的电脉冲方法以及哺乳动物细胞中常用的转染法导入外源基因,并对两种方法进行比较。电脉冲最佳 导入条件为:脉冲电压为300V/cm,脉冲次数为5,外源DNA质量浓度为20mg/L;转染法的最佳操作时间为受精后50min。应用上述两种方法进 行批量转基因鱼实验,得到外源基因的导入率分别为20%～28%和60%～70%。实验表明利用转染试剂jetPEI处理单细胞期的大菱鲆受精卵能得到较 高的孵化率及较高的转基因效率。 |
| 关键词: 转基因效率 电脉冲精子介导法 转染 大菱鲆(Scophthat musmaximus) |
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| 基金项目:国家863计划资助项目(2004AA628110);国家973计划(2004CB117402);青岛市科技局资助项目 |
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| Using electroporation and transfection methods to transfer foreign gene into Turbot (Scophthat musmaximus) |
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| Abstract: |
| This paper explores the relative efficiency of gene transfer into Turbot (Scophthat musmaximus) zygotes by electroporation and transfection reagent. The electroporation conditions(field strength of 300 V/cm, 5 pluse and foreign DNA concentration of 20 mg/L) were applied for mass gene transfer. Transfection experiments were performed at the one-cell stage (within 50 min postspawning) of fertilized fish eggs. Efficiency rates of foreign gene in the mass gene transfer were about 20%～28% and 60%～70% for electroporation and transfection methods, respectively. This study demonstrates that treating the fish zygotes with the DNA/jetPEI complex at the one-cell stage exhibits a higher gene transfer efficiency in fish. |
| Key words: transgenic efficiency electroporation transfection Scophthat musmaximus |
