Agroinfection as a rapid method for propagating Cowpea mosaic virus-based constructs
Agroinfection as a rapid method for propagating Cowpea mosaic virus-based constructs
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DOI：
10.1016/s0166-0934(02)00121-0
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发表时间：
2002-09-01
影响因子：
3.1
通讯作者：
Lomonossoff, GP
中科院分区：
文献类型：
--
作者：
Liu, L;Lomonossoff, GP
为了提高用基于Con;pea mosaic virus（CPMV）的构建体感染的效率，构建了适合农杆菌感染的克隆。将RNA-1和RNA-2的全长拷贝插入花椰菜花叶病毒（CaMV）35 S启动子和nos终止子的序列之间，并引入根癌农杆菌质粒pBINPLUS中。用含有基于RNA-1和RNA-2的质粒的混合物的细菌悬浮液浸润本氏烟草或豇豆（豇豆）的叶导致植物发展出典型的CPMV症状。为了证实该方法用于基于CPMV的载体的效用，基于RNA-2的GFP构建体被改造用于农杆菌感染。N.用含有该构建体和RNA-1质粒的农杆菌混合物处理本塞姆氏菌叶，导致高水平的GFP表达。结果表明，农杆菌感染是一种合适的方法，用于繁殖的CPMV为基础的衍生物。(C)2002 Elsevier Science B. V.保留所有权利。
To increase the efficiency of infections with Con;pea mosaic virus (CPMV)-based constructs, clones suitable for agroinfection were constructed. Full-length copies of RNA-1 and RNA-2 were inserted between the sequence of a Cauliflower mosaic virus (CaMV) 35S promoter and a nos terminator and were introduced into the Agrobacterium tumefaciens plasmid, pBINPLUS. Infiltration of leaves of either Nicotiana benthamiana or cowpea (Vigna unguiculata) with a bacterial suspension containing a mixture of the RNA-1- and RNA-2-based plasmids resulted in the plants developing typical CPMV symptoms. To confirm the utility of this approach for use with CPMV-based vectors, a GFP construct based on RNA-2 was adapted for agroinfection. Infiltration of N. benthamiana leaves with a mixture of Agrobacteria containing this construct and the RNA-1 plasmid resulted in high levels of GFP expression. The results demonstrate that agroinfection is a suitable method for the propagation of CPMV-based derivatives. (C) 2002 Elsevier Science B.V. All rights reserved.