摘要
以烟草叶片为试材,用转入植物表达载体pBI121的根癌农杆菌EHA105进行遗传转化。通过分析菌液不同浓度、侵染时间、乙酰丁香酮浓度及共培养时间对GUS基因表达的影响,研究其瞬时表达的最佳条件。结果显示:用OD600值为0.6的农杆菌侵染6min,在培养基中添加120μmol.L-1的乙酰丁香酮,共培养2d,能够得到高效的GUS基因瞬时表达。
An experiment was conducted to investigate the transient expression mediated by Agrobacterium using the leaves of tobacco （Nicotiana tabacum L.） as explants and the Agrobacterium tumefaciens EHAl05 containing the plant expression vector pBI121 for genetic transformation.Effects of A.tumefaciens concentration,infection time,acetosyringone concentration,and co-culture length on the transient expression of GUS gene were analysed to determine the optimal transformation condition.A higher transient expression level of GUS gene could be obtained as OD600 value of A.tumefaciens for infiltration 0.6,infiltrating for 6 min,adding 120 μmol·L^-1 acetosyringone to co-culture medium,and co-culture for 2 days in darkness.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2010年第9期110-112,共3页
Journal of Northeast Forestry University
基金
国家自然科学基金(30972387)资助
关键词
烟草
农杆菌转化
GUS基因
瞬时表达
Tobacco
Agrobacterium-mediated transformation
GUS gene
Transient expression
