Development of Arabidopsis Proteome Chip
Funded by NSF-2010 program
MCB-031514; 9/15/03-8/31/05
DBI-0519853; 11/1/05-10/31/07
DBI-0723722 & DBI-1042344; 2/15/08-8/31/12
PI: Mike Snyder; Co-PIs: S. P Dinesh-Kumar & Mark Gerstein
MCB-031514; 9/15/03-8/31/05
DBI-0519853; 11/1/05-10/31/07
DBI-0723722 & DBI-1042344; 2/15/08-8/31/12
PI: Mike Snyder; Co-PIs: S. P Dinesh-Kumar & Mark Gerstein
To develop a platform for large-scale analysis of plant proteins, we have generated 17400 Arabidopsis ORFs as tandem affinity purification (TAP) tag fusions. This Arabidopsis thaliana expression collection (ATEC) was used to produce recombinant Arabidopsis proteins using an optimized medium-throughput plant-based expression system. Purified recombinant proteins were printed to produce a high-density Arabidopsis protein microarrays (Popescu, et al., 2007 and 2009; Lee et al., 2011). We have used these arrays to perform protein-protein interaction studies (Popescu et al., 2007; Lee et al., 2011) and to identify targets of MAPKs (Popescu, et al., 2009).
Following materials are available from ABRC (http://abrc.osu.edu)
Materials and Protocols
References
Popescu, S. C, Popescu, G. V., Bachan, S., Zhang, Z., Seay, M., Gerstein, M., Snyder, M., and Dinesh-Kumar, S.P. (2007) Differential binding of calmodulin related proteins to their targets revealed using high-density Arabidopsis protein microarrays. Proc. Natl. Acad. Sci. 104:4730-4735. [Abstract]
Popescu, S. C, Snyder, M., and Dinesh-Kumar, S.P. (2007) Arabidopsis protein microarrays for the high-throughput identification of protein-protein interactions. Plant Signal Behav 2:416-420. [Abstract]
Popescu, S. C, Popescu, G. V., Bachan, S., Zhang, Z., Gerstein, M., Snyder, M., and Dinesh-Kumar, S.P. (2009) MAPK target networks in Arabidopsis thaliana revealed using functional protein microarrays. Genes & Development 2008 Dec 18. [Abstract]
Popescu S. C., Popescu, G.V., Snyder, M., and Dinesh-Kumar, S. P. (2009) Integrated analyses of co-expressed MAP kinase substrates in Arabidopsis thaliana. Plant Signal Behav. 4:524-527. [Abstract]
Lee, H. Y., Bowen, C., Popescu, G., Kang, H., Kato, N., Ma, S., Dinesh-Kumar, S.P., Snyder, M., and Popescu, S. C. (2011) Arabidopsis RTNLB1 and RTNLB2 reticulon-like proteins regulate intracellular trafficking and activity of the FLS2 immune receptor. Plant Cell (on line).
- pYL436 and pLIC-C-TAP expression vectors
- 17400 Arabidopsis ORFs fused to TAP tag in the ATEC collection
- Arabidopsis protein chip containing first 5000 proteins
- Arabidopsis protein chip containing additional 10000 proteins (will be available soon)
Materials and Protocols
- Vectors –pYL436 (Gateway-C-TAP) & pLIC-C-TAP (download)
- ATEC clones – Excel file (download)
- LIC cloning protocol (download)
- Protein expression protocol (download)
- Protein purification protocol (download)
- Protein printing and probing (download)
References
Popescu, S. C, Popescu, G. V., Bachan, S., Zhang, Z., Seay, M., Gerstein, M., Snyder, M., and Dinesh-Kumar, S.P. (2007) Differential binding of calmodulin related proteins to their targets revealed using high-density Arabidopsis protein microarrays. Proc. Natl. Acad. Sci. 104:4730-4735. [Abstract]
Popescu, S. C, Snyder, M., and Dinesh-Kumar, S.P. (2007) Arabidopsis protein microarrays for the high-throughput identification of protein-protein interactions. Plant Signal Behav 2:416-420. [Abstract]
Popescu, S. C, Popescu, G. V., Bachan, S., Zhang, Z., Gerstein, M., Snyder, M., and Dinesh-Kumar, S.P. (2009) MAPK target networks in Arabidopsis thaliana revealed using functional protein microarrays. Genes & Development 2008 Dec 18. [Abstract]
Popescu S. C., Popescu, G.V., Snyder, M., and Dinesh-Kumar, S. P. (2009) Integrated analyses of co-expressed MAP kinase substrates in Arabidopsis thaliana. Plant Signal Behav. 4:524-527. [Abstract]
Lee, H. Y., Bowen, C., Popescu, G., Kang, H., Kato, N., Ma, S., Dinesh-Kumar, S.P., Snyder, M., and Popescu, S. C. (2011) Arabidopsis RTNLB1 and RTNLB2 reticulon-like proteins regulate intracellular trafficking and activity of the FLS2 immune receptor. Plant Cell (on line).