摘要
为了将基因枪转化方法应用于菊花的遗传改良和品种选育,以gus基因为报告基因,利用基因枪方法将其转入菊花外植体内,通过检测gus基因在菊花叶盘中的瞬时表达情况,分析了轰击距离、氦气压力、金粉用量、质粒DNA浓度、轰击后培养时间、轰击次数等参数对gus瞬时表达的影响。结果表明,以幼嫩叶片为受体材料,在射程为6 cm、氦气压力为7.584×106Pa,每枪金粉用量为400μg,质粒DNA用量为3μg,轰击2次的条件下,2 d后可检测gus基因的最佳表达活性和最高瞬时表达率。
In order to apply particle bombardment to the genetic improvement and variety breeding of chrysanthemum, gus gene was used as report gene, then transformed into the explants of chrysanthemum by particle bombardment. The transient expression of gus gene in the leaf discs of chrysanthemum were examined to analyze the effects of shoot distance, helium pressure, gold particle amount, plasmid DNA concentration, culture-resuming time and bombarding frequency on the expression of gus gene. The results showed that, in the conditions of 6 cm shoot distance, 7. 584 X 10^6Pa helium pressure, 400 μg gold particle per shoot, 3μg plasmid DNA and bombarding for 2 times, the most active transient expression of gus gene could be detected two days after par- ticle bombardment. The transient expression of gus gene was also effected by bombarding fre- quency.
出处
《河南农业科学》
CSCD
北大核心
2007年第11期81-84,共4页
Journal of Henan Agricultural Sciences
基金
教育部博士点基金项目(20040022022)
科技部国家转基因专项项目(JY03-B-28)
关键词
菊花
基因枪
GUS基因
瞬时表达
Chrysanthemum
Particle bombardment
Gus gene
Transient expression
