L.-Y. Lee
2012/4/5
Curriculum Vitae
LAN-YING LEE
Current address and telephone number:
Department of Biological Sciences
Hansen Life Sciences Research Building
Purdue University
West Lafayette, Indiana 47907-1392
TEL: 765) 494-4947
FAX: (765) 496-1496
email:
Education
Ph.D. 1991, Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
M.S. 1985, Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
B.S. 1982, Department of Medical Technology, School of Medicine, National Taiwan University, Taipei, R.O.C.
Employment
2005/July-present
Associate Research Scientist, Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
2004-present
Visiting Associate Research Fellow, Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan
1998/Dec.-2005/June
Assistant Research Scientist, Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
1996/Oct.-1998/Dec.
Research Associate, Department of Biological Sciences, Purdue University, West Lafayette, Indiana. Laboratory of Dr. Stanton B. Gelvin.
1992/Oct.-1996/Sept.
Post-Doctoral Research Associate, Department of Plant Pathology, University of California, Davis, California. Laboratory of Dr. Clarence I. Kado.
1992/Feb.-1992/Oct.
Post-Doctoral Research Associate, Department of Molecular Microbiology, Washington University, School of Medicine, St. Louis, Missouri. Laboratory of Dr. Douglas E. Berg.
Post-Doctoral Research Associate, Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut. Laboratory of Dr. Claire M. Berg.
1991-1992
Adjunct Associate Professor, Department and Institute of Veterinary Medicine, National Taiwan University, Taipei, R.O.C.
1990-1992
Associate Research Fellow, Molecular Biology Division, Development Center for Biotechnology, Taipei, R.O.C.
1987-1991
Ph.D. student in the Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
1986-1987
Research Scientist, Molecular Biology Division, Development Center for Biotechnology, Taipei, R.O.C.
1985-1986
Assistant Scientist, Process Development Division, Development Center for Biotechnology, Taipei, R.O. C.
1985-1988
Adjunct Instructor, Yuanpei Institute of Medical Technology, Hsing-Chu, Taiwan, R.O.C.
1983-1985
M.S. student in the Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
1982-1985
Teaching assistant, School of Medical Technology, National Yang-Ming Medical College, Taipei, R.O.C.
Teaching Experience
2010, December 7
Conducted a lecture and laboratory entitled “BiFC to image protein-protein interactions in plants” at the Bimolecular Fluorescence Complementation (BiFC) workshop held by Bindley Bioscience Center of Purdue University.
2008, July 7-11
Conducted lectures and a laboratory course entitled “Agrobacterium Biology and Plant Transformation” at the Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
2007, July 9-20
Conducted lectures and a laboratory course entitled “Agrobacterium Biology and Plant Transformation” at the Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
2006, November 26-December 9
Invited by the College of Lift Sciences, Zhejiang University, Hangzhou, China, as a specialist to assist in the establishment of Bimolecular Fluorescence Complementation to study protein-protein interactions in rice.
2006, June 19-23
Conducted lectures and a laboratory in a summer course entitled “Agrobacterium Biology and Plant Transformation” at the Institute of Botany, Academia Sinica, Taipei, Taiwan.
2005, May 30–June 10
Conducted a workshop entitled “Bimolecular Fluorescence Complementation”, at the Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan.
2004, June 21–July 1
Conducted lectures and a laboratory in a summer course entitled “Agrobacterium Biology and Plant Transformation”, at the Institute of Botany, Academia Sinica, Taipei, Taiwan.
1999 Spring
Lecturer in “Eukaryotic Transformation” course, Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
1997 Spring
Teaching Assistant in “Eukaryotic Transformation” course, Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
1992 Spring
Lecturer in “Basic Molecular Biology”, Department and Institute of Veterinary Medicine, National Taiwan University, Taipei, R.O.C.
1991
Lecturer of “Polymerase Chain Reaction (PCR) Workshop”, held by Ministry of Economics, Development Center for Biotechnology, Taipei, R.O.C.
1985 - 1988
Lecturer in “Haemotology” and “Clinical Virology” courses, Yuanpei Institute of Medical Technology, Hsing-Chu, Taiwan, R.O.C.
1982 - 1985
Teaching Assistant in “Microbiology”, “Haemotology”, “Toxicology”, and “Virology” courses and lecturer in “Animal Tissue Culture Laboratory” course, School of Medical Technology, National Yang-Ming Medical College, Taipei, R.O.C.
Research Experience
2011-present
The main focus of my research is on developing advanced applications of previously established bimolecular fluorescence complementation (BiFC) systems for detection of protein-protein interactions in plants. The first project is to screen a cDNA library against bait proteins directly in plants to identify interacting partners. The second project is to screen a peptide aptamer library to identify peptide aptamer members that phenocopy mutations in plants directly. My third current project is to investigate the subcellular localization of various Agrobacterium virulence effector proteins in Arabidopsis myosin mutant plants.
2009-2010
There were two main focuses of this research: In the first, I was in charge of a project of direct screening of a BiFC cDNA library against “bait” proteins in plants. This project had been carrying out at the Institute of Plant and Microbial Biology, Academia Sinica, Taiwan part of year 2009 and continued until 2010. The second focus was to investigate the movement of Agrobacterium VirE2 protein visually in living plant cells using bimolecular fluorescence complementation technologies that we have developed in our laboratory.
2007-2008
Research was mainly focused on the following areas:
1. In charge of the multicolor BiFC project and aptamer BiFC project: Both are advanced applications of previously established bimolecular fluorescence complementation systems for detection of protein interactions in plants.
2. In charge of two joint projects sponsored by Dow Agroscience: The first project was to optimize the protein expression by tobacco BY-2 cells. The second project was to optimize the plant transformation efficiency by modifying Agrobacterium strains.
3. Investigation of protein-protein interactions between Arabidopsis proteins and A. tumefaciens/rhizogenes proteins in planta.
4. Investigation of mechanisms by which over-expression of various Arabidopsis histone genes enhances plant transformation.
2005-2007
Research was mainly focused on the following areas: First, development and documentation of BiFC (Bimolecular Fluorescence Complementation) vectors; this work is part of a NSF 2010 multi-laboratory project. Using the technology we developed, we were able to investigate interactions of Arabidopsis and Agrobacterium virulence proteins in planta. Second, we worked with Dow AgroSciences Corp.on achieving major breakthroughs in Agrobacterium-based transformation efficiency leading to genotype independent transgenic production. Third, I assisted with a BRDC project: Enhancing Agrobacterium-mediated transformation by manipulating the plant genome: a multi-gene approach. s
2003-2005
Research was mainly focused on three areas: Understanding the role of plant genes (mainly importin a genes of Arabidopsis thaliana) and proteins in the process of Agrobacterium-mediated transformation; development of novel vectors that facilitate cloning of large DNA inserts and expressing cDNAs; and generation of Agrobacterium strains that carrying T-DNA on the chromosome. In order to characterize the biological significance of various importin a isoforms and interaction between them, we established a protein interaction detection system in planta, called bimolecular fluorescence complementation. The goal was not only to detect protein interactions in vivo but also to demonstrate the intracellular location of these interactions. In addition, I constructed a series of cloning vectors, binary vectors, and expression vectors containing “gateway” sites. These vectors are useful for the genetic manipulation of plants. In order to improve crop transformation efficiency, we are evaluating the effect of binary vector oriV and Agrobacterium strains on transformation efficiency and integrated transgene copy number. We constructed a series of vectors harboring different oriV (giving different copy numbers per cell) but with identical T-DNAs in various Agrobacterium strains to compare the transformation efficiency. One of the constructions places the T-DNA onto the chromosome of Agrobacterium strains to generate one copy of T-DNA per cell. This project involved sophisticated microbial genetic techniques.
1996-2002
Research was mainly focused on two areas: the elucidation of factors involved in tumorigenesis by Agrobacterium tumefaciens, and the development of efficient plant transformation systems to generate transgenic plants using Agrobacterium. Research projects included: A comparison of the infection efficiency of various Agrobacterium strains by quantifyting T-strand production in bacteria and correlating this with the early molecular events in plant cells (expression of T-strand genes) at various time periods after bacterial inoculation; elucidation of the mechanism of oncogenic suppression of Agrobacterium by the plasmid pSa by investigating tumorigenesis and both transient and stable transformation phenotypes in various plant species/assay systems; comparison of the mechanisms of oncogenic suppression by IncW and IncQ type plasmids using conjugation assays (fertility inhibition); investigation of the mechanism of oncogenic suppression by pSa using genetic approaches and a yeast two-hybrid system; use of Arabidopsis as a plant system to identify plant factors which contribute to Agrobacterium transformation. The generation of crown gall resistant plants was included in the final goal.
1992-1996
Research carried out at UC Davis was involved in investigating the oncogenic suppression of Agrobacterium by the IncW plasmid pSa, the regulatory function of the Osa protein in operon expression and conjugation inhibition, and mapping/genetic organization of the plasmid pSa.
Collaboration with Pasteur Institute in developing mucosal immunity in swine to prevent Shigella infection by expressing bacterial antigens in tomato plants and using them as a potential oral vaccine.
1991-1992
Two main projects were conducted at the Development Center for Biotechnology, Taipei, R.O.C. First, establishment of protein expression systems in E. coli. Manupulation of protein expression vectors using various promoters, such as the l phage pL promoter, the T7 polymerase promoter (pET), and lac and tac promoters, and putting in cis- or trans- controlling elements to enhance the activity. Second, development of a multivalent recombinant vaccine against swine vesicular disease.
1987-1991
My Ph.D. thesis examined the genetic organization of genes involved in carotenoid biosynthesis of the epiphytic bacteria Erwinia herbicola. Genetic, microbiological, and molecular biological approaches (such as transposon Tn5, Tn1000 transposition, genomic library construction, gene cloning, and mapping) were used to determine the genetic organization of the gene loci. Biochemical and bacterial genetic methods were also used for characterization of the gene products.
1985-1987
Worked in the Development Center for Biotechnology to develop the first genetic engineering product, Hepatitis B Virus Vaccine, in Taiwan. This work included construction of a yeast plasmid expressing Hepatitis B virus pre-surface and surface antigens, fed-batch fermentation control of recombinant Saccharomyces cerevisiae, immunological approaches, such as hybridoma techniques and ELISA, and biochemical assays for obtaining antibodies and for the detection of recombinant proteins.
1983-1985
M.S. thesis was focused on restriction fragment analyses of human herpes virus clinical isolates for subtyping. Established primary cell lines for isolating clinical virus strains, used both conventional virology methodology and molecular biology approaches. Laboratory of Dr. Wu-Ze Liu, Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
1982-1983
Characterization of heat shock proteins induced in animal tissue culture. Laboratory of Dr. Ling-Bai Ting, Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, R.O.C.
1981-1982
Undergraduate thesis focused on typing and subtyping clinical herpes simplex virus isolates using immunoneutralization/titration assays in animal tissue culture system and immunofluorescent microscopy. Laboratory of Dr. Chun-Nan Lee, Department of Medical Technology, Medical School, National Taiwan University, Taipei, R.O.C.
Professional Activity
Member of American Society for Microbiologists
Member of American Society for Plant Physiologists
Member of International Society for Molecular Plant-Microbe Interactions
Member of International Society for Plant Molecular Biologists
Member of Society for In Vitro Biology
Fellowship
1987-1990
Fellowship for Ph.D. program from Development Center for Biotechnology, Taipei, R.O.C.
Book chapters
Lee, L.-Y. 2006. Integration of genes into the chromosome of Agrobacterium tumefaciens C58, pp. 55-66. In K. Wang (ed.), Methods in Molecular Biology: Agrobacterium protocols. Vol. 44, Humana Press, Totowa, New Jersey.
Publications
Lee, L.Y., and Gelvin, S.B. 2012. Bimolecular fluorescence complementation for imaging protein-protein interactions in planta. In Methods in Molecular Biology: Plant Functional Genomics. (P. Springer, ed.). Humana Press, Totowa, NJ. In press.
Lee, L.-Y., Wu, F.-H., Hsu, C.-T., Shen, S.-C., Yeh, H.Y., Liao, D.-C., Fang, M.-J., Liu, N.-T., Gelvin, S.B., and Lin, C.-S. 2012. Screening a cDNA library for protein-protein interactions directly in planta. Submitted.
Oltmanns, H., Frame, B., Lee, L.-Y., Johnson, S., Li, B., Wang, K., and Gelvin, S.B. 2010. Generation of “backbone” free, low transgene copy plants by launching T-DNA from the Agrobacterium chromosome. Plant Physiol. 152: 1158-1166.
Wu, F.-H., Shen, S.-C., Lee, L.-Y., Lee, S.-H., Chan, M.-T., Lin, C.-S. 2009. Tape-Arabidopsis Sandwich - a simpler Arabidopsis protoplast isolation method. Plant Methods 5:16. Selected and evaluated by The Faculty of 1000 on Jan. 4, 2010.
Hodges, L. D., Lee, L.-Y., McNett, H., Gelvin, S. B., and Ream, W. 2009. Agrobacterium rhizogenes GALLS gene encodes two secreted proteins required for gene transfer to plants. J. Bacteriol. 191: 365-374.
Tenea, G. N., Spantzel, J., Lee, L.-Y., Zhu, Y., Lin, K., Johnson, S., and Gelvin, S. B. 2009. Overexpression of several Arabidopsis histone genes increases Agrobacterium-mediated transformation and transgene expression in plant cells. Plant Cell 21:3350-3367.
Lee, L.-Y., Fang, M.-J., Kuang, L.-Y. and Gelvin, S. B. 2008. Vectors for multi-color bimolecular fluorescence complementation to investigate protein-protein interactions in living plant cells. Plant Methods. 4:24.
Bhattacharjee, S., Lee, L.-Y., Cao, H., Veena, and Gelvin, S.B. 2008. AtImpa-4, an Arabidopsis importin a isoform, is preferentially involved in Agrobacterium-mediated plant transformation. Plant Cell 20: 2661-2680. Selected and evaluated by The Faculty of 1000.
Lee, L.-Y. and Gelvin, S. B. 2008. T-DNA Binary Vectors and Systems. Plant Physiology 146:325-332.
Lee, L.-Y., Kononov, M. E., Bassuner, B., Frame, B. R., Wang, K., and Gelvin, S. B. 2007. Novel plant transformation vectors containing the superpromoter. Plant Physiology 145:1294-1300.
Citovsky, V.*, Lee, L.-Y.*, Vyas, S., Glick, E., Chen, M.-H., Babb, K., Vainstein, A., Gafni, Y., Gelvin, S. B., and Tzfira, T. 2006. Subcellular localization of interacting proteins by bimolecular fluorescence complementation in planta. J. Mol. Bio. 362:1120-1131.
*These authors contributed equally to this work.
Gaspar, Y.M., Nam, J., Schultz, C.J., Lee, L.-Y., Gilson, P., Gelvin, S.B., and Bacic, A. 2004. Characterization of the Arabidopsis lysine-rich arabinogalactan-protein AtAGP17 mutant (rat1) that results in a decreased efficiency of Agrobacterium transformation. Plant Physiol. 135: 2162-2171.