番茄转录因子LeMADS-MC正反义植物表达载体的构建

Construction of Tomato Transcription Factor LeMADS-MC Sense and Antisense Plant Expression Vector

为进一步阐明转录因子家族MADS-Box对番茄果实成熟的调控途径,本研究利用聚合酶链式反应技术(PCR)从番茄(Solanum lycopersicum)cDNA中克隆LeMADS-MC基因和LeMADS-antiMC基因核心片段,将这2个基因通过酶切分别正向连接到pCAMBIA1300-221载体和反向连接到pCXSN载体中,并用冻融法将重组载体导入农杆菌中。获得的重组载体分别通过PCR及酶切鉴定符合预期结果且测序结果与NCBI同源性高达100%。结果成功构建了适合于番茄农杆菌遗传转化的植物表达载体,为下一步通过LeMADS-MC基因研究果实成熟衰老机理奠定了物质基础。

To reveal the pathway of MADS-Box transcription factor family regulating tomato fruit ripening, the research obtained tomato （Solanum lycopersicum） LeMADS-MC gene and LeMADS-antiMC gene by the polymerase chain reaction technique （PCR）. The genes were connected to the plasmid pCAMBIA1300-221 and pCXSN, and then transformed into Agrobacterium by Freeze-thaw method. Vectors were identified by PCR and enzyme digestion in the research and the sequence result was homology to NCBI database up to 100%. The results indicated that tomato expression vectors pCAMBIA1300-221-MC and pCXSN-antiMC was constructed successfully. It would provide base for genetic research on fruit ripening and senescence through LeMADS-MC transgenic fruit.