抗冷冻蛋白基因(afp)转化大豆的研究

目的:通过农杆菌介导法遗传转化大豆。方法:通过热激法将质粒pCAAFP66导入根癌农杆菌菌株EHA105中获得含有抗冷冻蛋白基因(afp)及除草剂抗性筛选标记基因(bar)的农杆菌工程菌株;以大豆品种华春6号和马祖1号种子的下胚轴为外植体,经过农杆菌介导将抗冷冻蛋白基因导入大豆基因组中,在含有除草剂草丁膦(PPT)的培养基中筛选、并经过PCR鉴定获得大豆转化植株。结果:PPT的最佳筛选浓度为1.0mg/L,华春6号和马祖1号的阳性植株数分别为6株和2株,转化效率分别为3.70%和0.94%。结论:不同基因型大豆的转化率存在差异,抗冷冻蛋白基因成功遗传转化进大豆细胞中。

Transformation of Soybean with Antifreeze Protein Gene

Objective:With the help of Agrobacterium mediate to transfer Antifreeze Protein gene(afp) to soybean.Method:Through heat-shocking,the plasmid pCAAFP66 was transform into bacterial strains of Agrobacterium EHA105 to produce Agrobacterium project bacterial strains which contained Antifreeze Protein gene and herbicide resistence genes maker(bar).With the hypocotyls of soybean Huachun 6 and Mazu 1 as explants,Antifreeze Protein genes were led into the genomes of soybean through Agrobacterium mediate.In culture media which contained the herbicide glufosinate(PPT),the restructuring plants were sieved,and transformed soybean plants were initial confirmed through PCR evaluation.Result:The outcome showed that the best sieving density of PPT is 1.0mg/L,6 and 2 restructuring plants of Huachun 6 and Mazu 1 had been got,the transform efficiency are 3.70% and 0.94% respectively.Conclusion:There is some difference in the transform rate of soybeans with different types of genes,and the afp has been transferred into soybean cells.