中文摘要：

为了构建基于黄瓜花叶病毒（Cucumber mosaic virus,CMV）的外源蛋白表达载体,实验在获得CMV的LS株系（CMV-LS）农杆菌侵染性克隆（pCB301-R1,R2和R3）的基础上,将CMV-LS基因组RNA3的外壳蛋白（coat protein,CP）基因置换成绿色荧光蛋白（green fluorescent protein,GFP）基因,研究以CMV-LS为载体在本氏烟中表达GFP基因的情形。农杆菌浸润接种结果表明,以R1R2R3-gfp为载体在寄主中的GFP表达量远高于植物瞬时表达载体pCB301产生的GFP,番茄丛矮病毒基因沉默抑制子p19加入有助于CMV表达GFP,将移动蛋白（movement protein,MP）缺失C端的33个氨基酸残基而构建表达载体R1R2R3mpΔ33-gfp在寄主中的GFP含量大大增加。农杆菌菌液稀释接种的结果表明R1R2R3mpΔ33-gfp接种OD600值为0.01均能有效在本氏烟中表达产生GFP。

英文摘要：

In this study,Cucumber mosaic virus （CMV）,genomic RNAs of CMV LS strain were inserted into plant binary vector pCB301 respectively to obtain CMV-LS T-DNA infectious clones.In order to construct exogenous protein expression vector from CMV,coat protein （CP）gene in CMV genomic RNA3 was replaced with green fluorescent protein（GFP）and the seedlings of Nicotianabenthamiana were infiltrated with mixtures of A.tumefaciens containing plasmid pCB301-R1,R2 and R3-gfp to analyze the level of GFP expression.The results indicated that the accumulation of GFP expressed from R1R2R3-gfp was higher than that from transient expression vector pCB301-gfp and the addition of silencing suppressor protein p1 9 from Tomato bush stunt virus resulted in an increase in the amount of GFP.Deletion of the C-terminal 33 amino in CMV movement protein（MP）acids led to higher level of GFP expressed from vector R1R2 R3mpΔ33-gfp in the seedlings of N.benthamiana. The data of infiltration with bacterium suspension mixture of differnt dilution suggested that R1R2 R3mpΔ33-gfp vector could efficiently express GFP in N.benthamiana infected with infiltrated with mixtures of A.tumefaciens at optical density （OD600）of 0.01.