TY - JOUR
T1 - pSITE vectors for stable integration or transient expression of autofluorescent protein fusions in plants
T2 - Probing Nicotiana benthamiana-virus interactions
AU - Chakrabarty, Romit
AU - Banerjee, Rituparna
AU - Chung, Sang Min
AU - Farman, Mark
AU - Citovsky, Vitaly
AU - Hogenhout, Saskia A.
AU - Tzfira, Tzvi
AU - Goodin, Michael
PY - 2007/7
Y1 - 2007/7
N2 - Plant functional proteomics research is increasingly dependent upon vectors that facilitate high-throughput gene cloning and expression of fusions to autofluorescent proteins. Here, we describe the pSITE family of plasmids, a new set of Agrobacterium binary vectors, suitable for the stable integration or transient expression of various autofluorescent protein fusions in plant cells. The pSITE vectors permit single-step Gateway-mediated recombination cloning for construction of binary vectors that can be used directly in transient expression studies or for the selection of transgenic plants on media containing kanamycin. These vectors can be used to express native proteins or fusions to monmeric red fluorescent protein or the enhanced green fluorescent protein and its cyan and yellow-shifted spectral variants. We have validated the vectors for use in transient expression assays and for the generation of transgenic plants. Additionally, we have generated markers for fluorescent highlighting of actin filaments, chromatin, endoplasmic reticulum, and nucleoli. Finally, we show that pSITE vectors can be used for targeted gene expression in virus-infected cells, which should facilitate high-throughput characterization of protein dynamics in host-virus interactions.
AB - Plant functional proteomics research is increasingly dependent upon vectors that facilitate high-throughput gene cloning and expression of fusions to autofluorescent proteins. Here, we describe the pSITE family of plasmids, a new set of Agrobacterium binary vectors, suitable for the stable integration or transient expression of various autofluorescent protein fusions in plant cells. The pSITE vectors permit single-step Gateway-mediated recombination cloning for construction of binary vectors that can be used directly in transient expression studies or for the selection of transgenic plants on media containing kanamycin. These vectors can be used to express native proteins or fusions to monmeric red fluorescent protein or the enhanced green fluorescent protein and its cyan and yellow-shifted spectral variants. We have validated the vectors for use in transient expression assays and for the generation of transgenic plants. Additionally, we have generated markers for fluorescent highlighting of actin filaments, chromatin, endoplasmic reticulum, and nucleoli. Finally, we show that pSITE vectors can be used for targeted gene expression in virus-infected cells, which should facilitate high-throughput characterization of protein dynamics in host-virus interactions.
KW - Agroinfiltration
KW - Confocal microscopy
KW - Nucleolus
KW - SYNV
KW - Transformation
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UR - http://www.scopus.com/inward/citedby.url?scp=34250669555&partnerID=8YFLogxK
U2 - 10.1094/MPMI-20-7-0740
DO - 10.1094/MPMI-20-7-0740
M3 - Article
C2 - 17601162
AN - SCOPUS:34250669555
SN - 0894-0282
VL - 20
SP - 740
EP - 750
JO - Molecular Plant-Microbe Interactions
JF - Molecular Plant-Microbe Interactions
IS - 7
ER -