| 农杆菌介导的怀山药叶片瞬时表达方法的建立 |
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| 作者单位： | ;1.河南师范大学生命科学学院;2.河南师范大学河南省绿色药材生物技术工程实验室;3.河南师范大学河南省高校道地中药材保育及利用工程技术研究中心 |
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| 摘 要： | 高效的瞬时表达体系是方便、快捷的研究怀山药基因启动子活性、基因功能和生产重组蛋白的新途径.本研究选取怀山药叶片为受体,以β-葡糖醛酸酶(GUS)和绿色荧光蛋白(GFP)的基因为报告基因,对共培养时间进行分析,得到二者瞬时表达的最初时间以及较佳观察时间,建立根癌农杆菌介导的怀山药瞬时表达技术体系.结果显示:当注射的菌液浓度OD600=0.6,共培养3d时,GUS基因和GFP基因开始有表达,二者适于观察的共培养时间分别为5d和4d.
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| 关 键 词： | 怀山药 叶片 瞬时表达 GUS GFP |
Establishment of Agrobacterium-mediated Transient Expression System in Leaf of Dioscorea Opposita |
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| Institution: | ,College of Life Science,Henan Normal University,Engineering Laboratory of Green Medicinal Material Biotechnology,Henan Province,Henan Normal University,Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs,University of Henan Province,Henan Normal University |
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| Abstract: | Transient expression system is a new way to conveniently and efficiently studying promoter activity,gene function and producing recombination protein in Dioscorea opposite.An Agrobacterium-mediated transient expression method with GUS and GFP as reporter was established in D.opposita.We analyzed the expression of GUS and GFP in different culture time after leaf infiltration and studied the dynamic of transient expression.The result showed that the initial expression of GUS and GFP could be obtained after three-day of co-culture and the highest expression could be acquired after five-day and four-day,respectively. |
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| Keywords: | Dioscorea opposita leaf transient expression GUS GFP |
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