摘要
利用RT-PCR扩增拟南芥螫合肽合成酶(AtPCS1)基因全序列.进一步构建AtPCS1的植物表达载体pB I 121-AtPCS1,转化农杆菌EHA105;然后用转化的农杆菌EHA105以叶盘法侵染苜蓿甘农一号叶片,在50mg/L Kan的筛选压下,经过约80～100d的筛选,获得57棵再生苗.随机取其中9棵再生苗进行PCR检测,其中6棵为阳性.初步鉴定表明AtPCS1基因已整合到苜蓿基因组中.
The full length of AtPCS1 gene was amplified by RT-PCR from Arabidopsis thaliana （ecotype Columbia）. Furthermore, AtPCS1 plant expression vector pB I 121-AtPCS1 was constructed and transfered into Agrobacterium EHA105 and AtPCS1 gene was transferred into alfalfa Gannong No.1 by leaf infection method. Transgenic alfalfa plants have been regenerated under the 50 mg/L Kan concentration pressure after 80-100 days. Six putative transgenic plants were screened by AtPCSl-specific PCR in nine randomly chosen samples. The primary rcsults showed that the AtPCS1 has been transferred into alfalfa.
出处
《兰州大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第6期33-38,共6页
Journal of Lanzhou University(Natural Sciences)
基金
甘肃省自然科学基金项目(3ZS042-B25-011)
兰州理工大学博士基金项目(SB08200602)
幅建省自然科学基金项目(D0410004)资助.
关键词
植物螯合肽合成酶
植物表达载体
苜蓿
转化
鉴定
phytochelatin synthase
plant expression vector
alfalfa
transformation
identify
