摘要
利用RT-PCR技术从紫花苜蓿中克隆出过氧化物酶基因,命名为MsPOD,并通过DNA重组技术将其连接到载体pBI121上,成功构建植物表达载体pBI-MsPOD;将表达载体转化到感受态农杆菌株GV3101中,利用花序浸泡法获得具有卡纳抗性的转基因拟南芥植株。对其中3个再生植株进行PCR检测表明,目的基因已经成功整合到拟南芥基因组中,通过RT-PCR和GUS染色检测分析,初步证明目的基因在拟南芥中成功表达。
A peroxidase gene was cloned from alfalfa, which was named as MsPOD. By DNA recombi- nation technology, MsPOD gene was inserted into plasmid pBIl21, and a plant expressing vector pBI-POD was constructed. Transgenic Arabidopsis thaliana plants were obtained by floral dip method with Agrobacterium tumefaciens GV3101 harboring pBI-POD plasmid and seeds of transgeinc Arabidopsis thaliana were selected on MS plates containing 50mg/L Kanamycin. PCR analysis of 3 kanamycin resistant plants showed that the target gene was integrated into the genomes of the Arabidopsis thaliana. RT-PCR and GUS staining analysis showed that MsPOD gene expressed in Arabidopsis thaliana.
出处
《中国草地学报》
CSCD
北大核心
2013年第3期6-11,共6页
Chinese Journal of Grassland
基金
"十二五"国家科技支撑计划课题(2011BAD17B01-01-3)
关键词
紫花苜蓿
过氧化物酶
拟南芥
转化
Alfalfa
Peroxidase
Arabidopsis thaliana
Transformation
作者简介
作者简介：张怡（1988-），女，山西运城人，内蒙古农业大学在读硕士研究生，研究方向为植物资源利用与保护，Email：oioiyiyi@163．com．
通讯作者，Email：jinhong915@163．com.
