【摘要】 目的:采用克隆的番茄E8启动子和夏侧金盏花虾青素合成相关酶基因Adketo构建果实特异表达载体,为利用植物基因工程进行虾青素生产提供新途径。方法:采用RT-PCR的方法,从夏侧金盏花中克隆到虾青素合成相关酶基因Adketo,同时以番茄基因组DNA为模板PCR扩增果实特异性启动子E8,克隆进T载体,菌液PCR初步鉴定后测序,序列分析。从T载体切下的E8启动子和Adketo基因经回收后同时插入到植物双元表达载体pBI121上,获得重组载体,然后进行菌液PCR检测和酶切鉴定。结果:测序结果分析显示获得正确的E8和Adketo基因序列。重组质粒经菌液PCR检测及酶切鉴定均获得预期大小条带。结论:该实验成功构建了番茄果实特异性E8启动子驱动虾青素合成相关酶基因Adketo的植物表达载体。更多还原

【Abstract】 Objective:To construct the plant expression vector including Adketo gene from Adonis aestivalis and E8 promotor,and provide a new way for astaxanthin production by plant genetic engineering.Methods:The Adketo gene was clonied from Adonis aestivalis by RT-PCR and the 1.1 kb tomato E8 gene was amplified by PCR.then cloned into T vector.After the preliminary identification by Microbial PCR,the recombinant T-vectors were subjected to sequence analysis.Cut the E8 promoter and Adketo gene and inserted into the plant binary expression vector pBI121 to obtain the recombinant expression vector.And then to identify the vector by microbial PCR detection and enzyme digestion.Results:Sequencing results analysis showed the right E8 and Adketo gene sequences,the identification by microbial PCR and digestion with restriction enzymes proved that the recombinant vector had the inserts with expected length of target fragments.Conclusion:The plant expression vector containing Adketo gene driven by fruit specific promoter E8 was successfully constructed.更多还原