摘要
本研究通过简单检测DsRed2基因的表型就能快速筛出含有目的基因的成熟籽粒。DsRed2是一种在激发光下呈红色的荧光蛋白,根据在GenBank中获得的DsRed2及其特异性启动子Ltp2的基因序列,设计特异性引物,并构建筛选标记为Bar基因的植物表达载体pCAMBIA3300-Ltp2-DsRed2,通过农杆菌介导法转入玉米品种郑单958中。经除草剂筛选获得210株抗性植株,PCR检测得到阳性植株80株,对PCR检测呈阳性的植株进行试纸条检测,结果表明红色荧光蛋白基因DsRed2已成功整合到玉米基因组中。通过标记基因的快速检测,从而减少后代筛选的工作量和降低制种成本,为转基因玉米新品种的筛选提供直观有效的筛选标记。
The ai m of this study was to screen the mature seeds of the target genes by simple detecting the phenotype of DsRed2 gene. DsRed2 was a kind of red fluorescent protein under excitation light. According to the sequence of DsRed2 gene and seed coat tspecific promoter Ltp2 in GenBank, specific primers were designed, the plant expression vector pCAMB1A3300-Ltp2-DsRed2 which marker for Bar gene was selected and constructed, and transferred into maize variety Zhengdan958 by Agrobacterium mediated method. 210 plants were obtained by herbicide resistance screening, 80 plants with positive strains after PCR detection, at the same time, the PCR positive plants were tested in strip. The results showed that the red fluorescent protein gene DsRed2 has been successfully integrated into the maize genome. By the marker genes were rapidly detected to reduce the screening of offspring workload and production cost, which provided directly and effectively screening marker for the screening of new varieties of the transgenic maize.
出处
《分子植物育种》
CAS
CSCD
北大核心
2017年第5期1718-1723,共6页
Molecular Plant Breeding
基金
吉林省科技厅产业技术创新战略联盟项目(20140309005NY)
国家支撑计划项目(2014BAD01B01)共同资助
