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Construction of Plant Virus Expression Vector pClYVV/CP/W and Expression of GFP
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作者 王振东 王晓华 +5 位作者 孟鹏 秦会权 郑新伟 刘芳普 薛立江 张敏 《Agricultural Science & Technology》 CAS 2009年第5期38-41,48,共5页
[ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein(GFP) with pCIYVV/CP/W, and to develop effective plat virus vector fo... [ Objective] The study was to report the construction of plant virus expression vector pCIYVV/CP/W and the expression of green fluorescent protein(GFP) with pCIYVV/CP/W, and to develop effective plat virus vector for plant bioreactor to produce useful protein. [ Method] A section of multiple cloning sites among NIb/CP genes in pCIYVV genome and deoxyribonucleotide polylinker of cleavage recognition sequence containing viral protease Nla were cloned with infectivity full-length cDNA of clover yellow vein virus (CIYVV), and pCIYVV/CP/W vector was constructed, GFP gene was inserted into pCIyVV/CP/W to construct the pCIYVV/CP/W/GFP vector. The transcription situation of recombinant virus clone was detected by RT-PCR, and targeted gene products expressed by recombinant virus clone were detected with western blot (WB). [Result] The broad bean seedling inoculated with pCIYVV/CP/W/GFP expressed the same symptom as wild type CIYVV, morbidity was of 100%, the result showed that recombinant virus clone pCIYVV/CP/W/GFP didn't suppress, insertion of foreign gene didn't destroy the open reading frame of pCIYVV/CP/W. Foreign gene can keep living in F, progeny virus genorne steadily, recombinant virus clone pCIYVV/CP/W/GFP could steadily express GFP in progeny virus at least.[ Conclusion] The useful plant virus vector was provided for useful protein expressing. 展开更多
关键词 plant virus expression vector pCIYVV/CP/W pCIYVV/CP/W/GFP CONSTRUCTION expression
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