Agrobacterium-mediated transformation of AtPCS1 into Medicago sativa.L

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 pBⅠ121-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 AtPCS1-specific PCR in nine randomly chosen samples.The primary results showed that the AtPCS1 has been transferred into alfalfa.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