ACC脱氨酶基因转化白兰瓜的初步研究

Primary study on transforming Cucumis melo var.I with ACC-deaminase gene

用带有 ACC脱氨酶基因和卡那霉素抗性基因 ( Npt ,作为报告基因 )的工程根癌农杆菌转化白兰瓜子叶 ,通过组织培养 ,得到具有卡那霉素 ( Km)抗性的再生小苗。经 Npt 报告基因的 PCR扩增、ACC脱氨酶基因的 Southern点杂交以及 ACC脱氨酶活性的生理生化检测可知 ,ACC脱氨酶基因已成功转入白兰瓜子叶再生小苗 ,而且不同植株的真叶中表现出不同水平的 ACC脱氨酶活性。

Cotyledons of Cucumis melo var.I were transformed with Agrobacterium tumefaciens (LBA4404) carrying ACC deaminase gene and Kanamycin resistant gene ( NptⅡ ,as report gene).Some regenerated Kanamycin resistant seedlings were obtained.Through PCR amplification of NptⅡ gene,southern dot blotting of ACC deaminase gene and physio chemical assays of the enzyme activity,it was showed that ACC deaminase gene was successfully transferred into several regenerated seedlings,and different activity of ACC deminase displayed in leaves of different seedlings.