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Agroinoculation as a Simple Way to Deliver a Tobacco Mosaic Virus- Based Expression Vector 被引量:7
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作者 贾洪革 庞永奇 方荣祥 《Acta Botanica Sinica》 CSCD 2003年第7期770-773,共4页
烟草花叶病毒(TMV)表达载体30B是一个目前广泛应用的植物病毒表达载体,但用其生产外源蛋白时,必须先将它体外转录成RNA,才能被用来接种宿主植物。由于RNA体外转录费用昂贵、操作复杂,因此限制了30B表达载体的进一步应用。针对这一不足,... 烟草花叶病毒(TMV)表达载体30B是一个目前广泛应用的植物病毒表达载体,但用其生产外源蛋白时,必须先将它体外转录成RNA,才能被用来接种宿主植物。由于RNA体外转录费用昂贵、操作复杂,因此限制了30B表达载体的进一步应用。针对这一不足,我们用农杆菌接种法(agroinoculation)接种该病毒载体,即将30B cDNA置于花椰菜花叶病毒(CaMV)的35S启动子和终止子之间,再将整个表达框架插入到农杆菌T-DNA的左边界和右边界之内,构建成质粒p35S-30B,将转入该质粒的农杆菌注射到植物的叶片中,30B cDNA随T-DNA进入植物细胞后,被转录成可自我复制的RNA形式,进而发生系统侵染。为了检测此接种方式的可行性,绿色荧光蛋白(GFP)报告基因被克隆到p35S-30B中,构建成p35S-30B∶∶GFP,用含有该质粒的农杆菌进行注射操作。证实该病毒载体可通过简便的农杆菌接种法侵染Nicotiana benthamiana,在被接种植物的系统叶中,GFP的表达量可占植物总可溶蛋白的5.2%。 展开更多
关键词 tobacco mosaic virus agroinoculation gene expression
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Molecular characterization and pathogenicity of an infectious cDNA clone of tomato brown rugose fruit virus
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作者 Ziyue Ma Hui Zhang +3 位作者 Ming Ding Zhongkai Zhang Xiuling Yang Xueping Zhou 《Phytopathology Research》 2021年第1期160-169,共10页
Tomato brown rugose fruit virus(ToBRFV)is a new member of the genus Tobamovirus,and has the potential to affect the production and marketability of tomatoes and peppers.In this study,we sequenced and analyzed the comp... Tomato brown rugose fruit virus(ToBRFV)is a new member of the genus Tobamovirus,and has the potential to affect the production and marketability of tomatoes and peppers.In this study,we sequenced and analyzed the complete genome of ToBRFV isolates from tomato plants showing mosaic and mottling symptoms in Yunnan Province of China.We constructed a full-length infectious cDNA clone of ToBRFV,which could induce systemic infection with typical symptoms in tomato,Nicotiana benthamiana,and N.tabacum cv.Samsun nn plants through Agrobacterium-mediated inoculation.Further experimental evidence demonstrated that the rod-shaped virions accumulating in agroinfiltrated plants are sap-transmissible.This is the first report on the construction of a biologically active,full-length infectious cDNA clone of ToBRFV.The system developed herein will facilitate further research on functions of ToBRFV-encoded proteins and plant-ToBRFV interactions through reverse genetic approaches. 展开更多
关键词 TOBAMOVIRUS Tomato brown rugose fruit virus Infectious clone agroinoculation
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