摘要
以番茄无菌苗的子叶为外植体,以抗卡那霉素的NPTⅡ作为选择标记基因,通过根癌农杆菌介导法,将甜蛋白Thaumatin Ⅱ基因导入番茄中,共获得10株独立的抗性转化株系。对抗性转化株系进行了PCR和Southern blotting鉴定,PCR检测得到了预期的特异条带,Southern blotting结果表明其中1个转化株系插入了2个拷贝的Thaumatin Ⅱ cDNA。以上试验结果证明Thaumatin Ⅱ基因已经整合到转基因番茄植株的基因组中,为最终获得在蛋白质水平表达Thaumatin Ⅱ的无选择标记的转基因番茄打下了基础。
Using sterile tomato cotyledons as explants and kanamycin-resistant NPT Ⅱ as a marker gene, the Thaumatin 17 gene which encodes a plant sweet protein from Thaumatococcus daniellii has been introduced into the tomato genome by Agrobacterium-mediated transformation. As a result, 10 independent tomato transgenic lines were acquired. The introduction of the foreign gene was confirmed by PCR for the expected specific bands. Furthermore, one transgenic line was shown by two copies of integration on Southern blotting analysis. Taken together, all the above results have indicated that the Thaumatin Ⅱ gene has integrated into the genome of tomato. This represents the first step to finally obtain marker-free transgenic tomatoes expressing plant sweet protein Thaumatin Ⅱ. transformation
出处
《浙江农业学报》
CSCD
北大核心
2010年第2期167-172,共6页
Acta Agriculturae Zhejiangensis
基金
浙江省农业科学院科技创新能力提升工程项目
作者简介
杨飞（1983-），男，江西九江人，主要从事番茄转基因技术研究。Tel：0571-86417305；E-mail：yanfeixyz_@126．com
通讯作者，杨悦俭，E-mail：hzyyj@yahoo．com．cn
