家稗Pdk基因叶片特异性表达载体的构建及农杆菌的导入

摘要/Abstract

摘要： 【研究目的】构建含家稗Pdk基因的叶片特异性表达载体；【方法】通过PstI和SalI单酶切分别从质粒pRGN及pUCm-Pdk上获得Rubisco小亚基启动子和Pdk基因，将其连入表达载体pCAMBI1301, 并通过冻融法将重组质粒导入根癌农杆菌EHA105；【结果】构建了由Rubisco小亚基启动子调控的Pdk基因植物表达载体p1301-Prbs-Pdk，选择标记基因为Hpt（潮霉素磷酸转移酶基因），经酶切和PCR鉴定，表达载体已成功导入农杆菌EHA105中；【结论】丙酮酸磷酸双激酶（PPDK）是C4光合途径中的关键光合酶，本实验中构建了含有rbcs启动子的适于单子叶植物转化的Pdk基因表达载体，为提高转基因植物光合效率奠定了基础。

关键词: 扶芳藤, 扶芳藤, 遗传多样性, RAPD鉴定, 类型划分

Abstract: 【OBJECTIVE】 The pyruvate-phosphate dikinase expressed effectively by utilizing leaf-specfic promoter of small subunits of rubisco; 【METHOD】The small subunits of rubisco and the gene of Pdk were digested by PstI and SalI from pRGN and pUCm-Pdk,and inserted into expression vector pCAMBI1301.Then it was transformed into EHA105 by direct entering way; 【RESULTS】The plant transformation vector p1301-Prbcs-PPDK was constructed in this experiment. The selective marker gene was Hpt(Hygomycin phosphate transferase). By digestion and PCR identification, The vector p1301-Prbcs-PPDK was transferred to Agrobacterium tumefaciens EHA105 ; 【CONCLUSION】 The pyruvate-phosphate dikinase is the key enzyme of C4 photosynthesis.The experiment constructed a expression vector suit to monocotyledon,which will provide an important experimental base for study on PPDK function..

中图分类号:

王金明,,赵明,丁在松,张斌,郭志江. 家稗Pdk基因叶片特异性表达载体的构建及农杆菌的导入[J]. 中国农学通报, 2007, 23(5): 63-63.

Wang Jinming,, Zhao Ming, Ding Zaisong, Zhang Bin, Guo Zhijiang. Construction of Plant Leaf-specific Expression Vector with PPDK of [Echinochloa crusgalli var frumentacea(Roxb.)W.F.Wight] and Transfer to Agrobacterium tumefaciens[J]. Chinese Agricultural Science Bulletin, 2007, 23(5): 63-63.

参考文献

[1] ap Rees T. Hill S A. Metabolic control analysis of plant metabolism. Plant Cell Environ,1994,17:587-599.
[2] Trevanion S J, Furbank R T, Ashton A R. NADP-malate dehydrogenase in the C4 plant Flaveria bidentis. Cosense suppression of activity in mesophyll and bundle-sheath cells and consequences for photosynthesis. Plant Physiol., 1997,113:1153-1165.
[3] Zhang L(张丽). Comparison of photosynthetic characters and their physiological mechanism in C3 and C4 crops. Dissertation of Master degree in China Agricultural University. (中国农业大学硕士学位论文), 2004(in Chinese).
[4] 刘巧泉,于恒秀,张文娟,等.水稻rbcs启动子控制的外源基因在转基因水稻中的特异性表达.植物生理与分子生物学学报(Journal of Plant Physiology and Molecular Biology). 2005, 31(3):247-253.
[5] Matsuoka M. Structure, genetic mapping, and expression of the gene for pyruvate, orthophosphate dikinase from maize. J Biol Chem, 1990, 265(28): 16772-16777.
[6] Sheen J. Molecular mechanisms underlying the differential expression of maize pyruvate, orthophosphate dikinase genes. Plant Cell, 1991,3(3):225-245.
[7] Ishimaru K, I chikawa H, Matsuoka M,Ohsugi R. Analysis of a C4 maize pyruvate, orthophosphate dikinase expressed in C3 transgenic Arabidopsis plants.Plant Sci., 1997,129:57-64.
[8] Fukayama H, Tsuchida H, Agarie S, et al. Significant accumulation of C4-specific pyruvate, orthophosphate dikinase in a C3 plant, rice. Plant Physiology, 2001,127:1136-1146.
[9] Ku M S B, Cho D H, Li X, et al. Introduction of genes encoding C4 photosynthesis enzymes into rice plants physiological consequences Rice Biotechnology. Improving Yield, Stress Tolerance and Grain Quality, 2001, 236:100-116.
[10] Morris S H, Adley C C. Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods[J]. Trends Biotechnol, 2001,19(2):43.
[11] Kuiper H A, Kleter G A, Noteborn H P, et al. Assessment of the food safety issues related to genetically modified foods[J]. Plant J, 2001,27(6):503.

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