农杆菌介导的大豆反义fad2-1基因转化烟草研究  

Transfer of Antisense Fatty Acid Desaturase Gene from Soybean to Tobacco Plantlets (Nicotiana tabaccu) via Agrobacterium-mediated Transformation

作　　者：范红军[1] 李敏[1] 周延清[1] 楚素霞[1] 邢延豪[1] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007

出　　处：《河南师范大学学报（自然科学版）》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)

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.

关 键 词：烟草 反义油酸脱饱和酶基因 根癌农杆菌 遗传转化 

分 类 号：Q788[生物学—分子生物学]