摘要
以烟草无菌苗叶片为外植体,通过根癌农杆菌介导法,将大豆反义油酸脱饱和酶基因导入烟草中,经梯度卡那霉素筛选,获得可在含75mg/L卡那霉素选择生根培养基上再生的抗性植株75株,其中67株抗卡那霉素植株的PCR分析扩增出目的片段;RT-PCR分析表明该基因在烟草中表达.结果表明大豆反义油酸脱饱和酶基因在烟草基因组中整合和表达.这为大豆反义油酸脱饱和酶基因转入大豆和表达奠定基础,也为烟草基因工程育种提供了依据.
The antisense Δ^12-fatty acid desaturase gene cloned from soybean genome is introduced into tobacoo by Agrobacterium tumefaciens-mediated leaf disc transformation.75 kanamycin-resistant tobacco plantplets are regenerated in shoot induction culture and rooting culture supplemented with 75 mg/L Kanamycin. PCR amplification analysis showe that this gene is successfully integrated into the genomes of 67 kanamycin-resistant tobacco plants. RT-PCR detection reveals that the gene is expressed in transformed tabacoo plantlets. This has laid the foundations for the transfer and expression of fad-1 gene to soybean and tabacco gene engineering breeding.
出处
《河南师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第2期152-155,共4页
Journal of Henan Normal University(Natural Science Edition)
基金
国家863项目(2006AA100104-15)
河南省教育厅自然科学基础研究项目(2008A208018)
关键词
烟草
反义油酸脱饱和酶基因
根癌农杆菌
遗传转化
tobacoo（Nicotiana tobacum.cv. Xanthi）
antisense oleic acid desaturase gene
Agrobacterium tumefaciens
genetic transformation.
