This Year 2 Progress Report is Cumulative for the Entire Term of the Grant

Names and Positions of People Hired
Research Progress
Encountered Problems

A. Names and Positions of People Hired

B. Publications using basic/early versions of BiFC constructs
  1. 1. Tzfira, T., Vaidya, M., & Citovsky, V. (2004) . Nature 431, 87-92.
    2. Loyter, A., Rosenbluh, J., Zakai, N., Li, J., Kozlovsky, S.V., Tzfira, T., & Citovsky, V. (2005) The plant VirE2 interacting protein 1. A molecular link between the Agrobacterium T-complex and the host cell chromatin? Plant Physiol. 138, 1318-1321.
    3. Lacroix, B., Vaidya, M., Tzfira, T., & Citovsky, V. (2005) The VirE3 protein of Agrobacterium mimics a host cell function required for plant genetic transformation. EMBO J. 24, 428-437.
    4. Li, J., Krichevsky, A., Vaidya, M., Tzfira, T., & Citovsky, V. (2005) Uncoupling of the functions of the Arabidopsis VIP1 protein in transient and stable plant genetic transformation by Agrobacterium. Proc. Natl. Acad. Sci. USA 102, 5733-5738.
    5. Lacroix, B., Tzfira, T., Vainstein, A., & Citovsky, V. (2005) A case of promiscuity: Agrobacterium’s endless search for partners. Trends Genet. 21, in press.
    6. Gelvin, S.B. (2005) Gene exchange by design. Nature 433, 583-584.
    7. Gelvin, S.B. (2005) Viral-mediated plant transformation gets a boost. Nature Biotechnol. 23, 684-685.
    8. Tzfira, T., Tian, G.W., Lacroix, B., Vyas, S., Li, J., Leitner-Dagan, Y., Krichevsky, A., Taylor, T., Vainstein, A., & Citovsky, V. (2005) pSAT vectors: a modular series of plasmids for fluorescent protein tagging and expression of multiple genes in plants. Plant Mol. Biol. 57,
    9. Chung, S.-M., Frankman, E.L., & Tzfira, T. (2005) A versatile vector system for multiple gene expression in plants. Trends Plant Sci. 10, 357-361.

    Publications using current versions of BiFC constructs
    10. Krichevsky, A., Kozlovsky, S.V., Gafni, Y., & Citovsky, V. (2006) Nuclear import and export of plant viral proteins and genomes. Mol. Plant Pathol. 7, 131-146. COVER ARTICLE
    11. Citovsky, V., Lee, L.Y., Vyas, S., Glick, E., Chen, M.H., Vainstein, A., Gafni, Y., Gelvin, S.B., & Tzfira, T. (2006) Identification of subcellular localization of interacting proteins by bimolecular fluorescence complementation in planta. J. Mol. Biol. 362, 1120-1131.
    12. Gelvin, S.B. (2007) Function of host proteins in the Agrobacterium-mediated plant transformation process. In Agrobacterium, from biology to biotechnology (T. Tzfira and V. Citovsky, eds.). Kluwer Academic Publishers, Dordrecht, The Netherlands. In press.
    13. Bhattacharjee, S., Cao, H., Lee, L.-Y., Veena, and Gelvin, S.B. 2006. AtImpa-4, an Arabidopsis importin ? isoform, is preferentially involved in Agrobacterium-mediated plant transformation. Submitted.
C. Research Progress

During the first year of the grant, we performed two lines of concurrent experiments: proof of concept of BiFC in plants using basic, early versions of the vectors, and converting these vectors to the optimized BiFC system described in the original proposal.

D. Encountered Problems

We have found that in electroporated tobacco BY-2 protoplasts, signal from  slowly maturing dsRed protein gives significant fluorescence in the YFP channel.  We have overcome this problem by using mRFP instead of dsRed.