| RRC ID |
3989
|
| Author |
Ogawa Y, Dansako T, Yano K, Sakurai N, Suzuki H, Aoki K, Noji M, Saito K, Shibata D.
|
| Title |
Efficient and high-throughput vector construction and Agrobacterium-mediated transformation of Arabidopsis thaliana suspension-cultured cells for functional genomics.
|
| Journal |
Plant Cell Physiol
|
| Abstract |
We established a large-scale, high-throughput protocol to construct Arabidopsis thaliana suspension-cultured cell lines, each of which carries a single transgene, using Agrobacterium-mediated transformation. We took advantage of RIKEN Arabidopsis full-length (RAFL) cDNA clones and the Gateway cloning system for high-throughput preparation of binary vectors carrying individual full-length cDNA sequences. Throughout all cloning steps, multiple-well plates were used to treat 96 samples simultaneously in a high-throughput manner. The optimal conditions for Agrobacterium-mediated transformation of 96 independent binary vector constructs were established to obtain transgenic cell lines efficiently. We evaluated the protocol by generating transgenic Arabidopsis T87 cell lines carrying individual 96 metabolism-related RAFL cDNA fragments, and showed that the protocol was useful for high-throughput and large-scale production of gain-of-function lines for functional genomics.
|
| Volume |
49(2)
|
| Pages |
242-50
|
| Published |
2008-2-1
|
| DOI |
10.1093/pcp/pcm181
|
| PII |
pcm181
|
| PMID |
18178967
|
| MeSH |
Arabidopsis / genetics*
Arabidopsis / metabolism*
Cell Line
Cells, Cultured
Gene Expression Profiling
Gene Expression Regulation, Plant
Genetic Vectors
Genome, Plant
Genomics / methods*
Plants, Genetically Modified
Rhizobium / physiology*
Transformation, Genetic
|
| IF |
4.062
|
| Times Cited |
33
|
|
WOS Category
|
PLANT SCIENCES
CELL BIOLOGY
|
| Resource |
| Arabidopsis / Cultured plant cells, genes |
rpc00008 |
