大叶补血草质膜Na^+/H^+逆向转运蛋白基因SOS1的克隆及对番茄的遗传转化

Cloning and Genetic Transformation of Tomato with SOS1 Gene from Limonium gmelinii

根据大叶补血草(Limonium gmelinii(Wildl.)Kuntze)SOS1基因序列(登录号:EU780458)设计特异性引物,通过RT-PCR方法从叶片中克隆得到质膜Na+/H+逆向转运蛋白基因LgSOS1,将该基因重组于质粒pCAMBIA1390的CaMV35S启动子下游,构建含LgSOS1基因植物表达载体pCAMBIA1390-LgSOS1。通过根癌农杆菌介导法转化番茄(Lycopersicon esculen-tum),在1.2 mg/L6-BA、0.2 mg/L IAA、20 mg/L潮霉素和200 mg/L羧苄青霉素的MS培养基上进行选择培养,从抗性愈伤组织中获得再生植株。经PCR和RT-PCR检测,初步证实LgSOS1基因已整合至番茄的基因组中。

A plasma membrane-typed Na＋/H＋ antiporter gene SOS1 has been separated from halophyte Limonium gmelinii（Wildl.）Kuntze by RT-PCR.Expression cassettes including a CaMV35S promoter,the Na＋/H＋ antiporter gene LgSOS1,and NOS poly A sequence were cloned into the vector pCAMBIA1390,which was transferred into tomato by Agrobacterium-mediated method.Transgenic tomatoes were regenerated and selected on MS basal medium supplemented with 1.2 mg/L 6-BA,0.2 mg/L IAA,20 mg/L Hygromycin and 200 mg/L carbenicillin.Some Hygromycin-B-resistant transgenic tomatoes were obtained and confirmed by PCR and RT-PCR.