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。
Abstract
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.
关键词
番茄成熟； LeMADS-MC； 克隆； 表达载体
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Key words
tomato ripening; LeMADS-MC; cloning; expression vector
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参考文献
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脚注
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