Improved expression of recombinant GFP using a replicating vector based on Beet curly top virus in leaf-disks and infiltrated Nicotiana benthamiana leaves.
Recombinant green fluorescent protein (GFP) with a molecular mass of 29 kDa was transiently expressed in Agrobacterium-inoculated leaf-disks prepared from Nicotiana benthamiana plants. Expression of GFP from the Cauliflower mosaic virus (CaMV) 35 S promoter within a replicating vector based on the geminivirus Beet curly top virus (BCTV) was more than 3 times higher than from a control, non-replicating vector. Use of the Cassava vein mosaic virus (CsVMV) promoter in the BCTV replicating vector increased the expression of recombinant GFP 320% at the transcript level, compared to use of the control CaMV 35 S promoter. Expression of recombinant GFP from Agrobacterium-inoculated leaf-disks of N. benthamiana was further enhanced up to 240% in the presence of post-transcriptional gene silencing suppressor p19.
Kim
Kyung Il
KI
Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University, Suwon 449-701, Korea.
Sunter
Garry
G
Bisaro
David M
DM
Chung
In Sik
IS
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004261:DNA Replication; D017791:Geminiviridae / Q000235:genetics*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D014026:Nicotiana / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D003546:Cysteine Endopeptidases; C072775:HC-Pro protein, potyvirus