PMI基因作为选择标记的植物表达载体构建及其在雪柑转基因中的应用  

Construction of Plant Expression Vectors with PMI Gene as Selection Marker and Their Utilization in Transformation of Citrus sinensis

作　　者：曾黎辉[1] 徐海峰[1] 王会全[1] 吴少华[1] 朱艺萱[1] 

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《农业生物技术学报》2008年第5期858-864,共7页Journal of Agricultural Biotechnology

基　　金：福建省自然科学基金(No.B0510018);教育部留学回国人员科研启动基金资助

摘　　要：以大肠杆菌(Escherichia coli)6-磷酸甘露糖异构酶(6-phosphomannose isomerase,PMI)基因替换植物表达载体pCAMBIA1301中的hpt基因以及pBI121中的gus基因,构建了以PMI基因为选择标记基因的植物表达载体pCAMBIA1301PMI和pBIPMI,并导入根癌农杆菌(Agrobacterium tumefaciens)EHA105中。研究了两种表达载体对雪柑(Citrus sinensis L.Osbeck)上胚轴的转化,在培养基附加25g/L甘露糖和5g/L蔗糖为碳源的选择压力下,pCAMBIA1301PMI的转化率为27.7%,pBIPMI的转化率为12.7%,对再生植株用氯酚红和PCR检测证实了PMI基因的导入,建立了以PMI/甘露糖为选择系统的雪柑转基因体系。The plant expression vectors pCAMBIA1301PMI and pBIPMI were constructed by substituting Escherichia coil phosphomannose- isomerase （PMI） gene for hpt gene of pCAMBIA1301 and gus gene of pBI121. Epicocyl explants of Xuegan sweet orange （Citrus sinensis L.Osbeck） were inoculated with Agrobacterium-tumefaciens EHA105- pCAMBIA1301PMI and EHA105- pBIPMI and subsequently selected on medium supplemented with a combination of 25 g/L mannose and 5 g/L sucrose as a carbon source. The transformation efficiency rate was 27.7% when transformed by pCAMBIA1301PMI and 12.7% by pBIPMI. Genetic transformation was confirmed by chlorophenol red assay and PCR, indicating that a new method for obtaining transgenic Xuegan plants was developed using PMI/mannose selection system.

关 键 词：6-磷酸甘露糖异构酶(PMI) 基因 甘露糖 雪柑 转基因 

分 类 号：S188[农业科学—农业基础科学]