Assistant Construction

Eugene N. Zaitsev

Section on Neural Development and

Plasticity PNRC, LCSN, NICHD, NIH

** ******* *****, **** **** Bethesda, MD 20892

Tel. 301-***-**** (home)

e-mail *************@*******.***

EDUCATION:

Ph.D. in Genetics, awarded by St. Petersburg State University. Research

work was carried out in the St. Petersburg Polytechnic State University,

St. Petersburg, Russia. Thesis:" Structural and Functional Study of RecA

like gene from Pseudomonas aeruginosa". Advisor: Professor V. Lanzov.

M.S. in Biophysics, St. Petersburg Polytechnic State University, St.

Petersburg, Russia. Thesis:"Study of the protein biosynthesis system of

yeast mitochondria". Advisor: Professor S. Bresler.

PROFESSIONAL EXPERIENCE:

2010- now Scientist, FinaBiosolutions, LLC. Gaithersburg, MD

2006- 2010 Volunteer, Section on Neural Development and Plasticity, and

Section on Biophysics, NICHD, NIH, Bethesda, MD

2000-2006 Research Fellow, Section on Neural Development and Plasticity,

NICHD, NIH, Bethesda, MD

Cell biology of BDNF trafficking and signal transduction in neurons.

1999-2000 Research Scientist in the Department of Psychiatry and Behavioral

Science, State University of New York at Stony Brook, NY.

Mapping mitotic TG3 phosphoepitope of human nucleolin. Study of processing

and intracellular trafficking of Alzheimer disease precursor protein in

vivo with fluorescent tag-protein chimeras.

1995-1999 Assistant Research Microbiologist at the University of California

at Davis, Davis, CA.

Characterization of the double-stranded DNA binding properties of RecA

protein by fluorescence methods. Identification of the DNA binding sites of

the RecA protein by mutational analysis. Inverse DNA strand exchange

reaction.

1991-1995 Postdoctoral Fellow at the University of California at Davis,

Davis, CA.

Biochemical and genetic analysis of E.coli RecA protein function.

Generation of N- and C-terminal truncations of the recA gene by PCR and

subsequent characterization of the mutant proteins.

1990-1991 Visiting Assistant Research Geneticist at the University of

California at Berkeley, Berkeley, CA.

Mapping new mutants in the RecA gene. Mapping the RecA protein epitope for

monoclonal antibody Mab4B4.

. 1987-1990 Senior Scientist at Molecular Genetics Laboratory at St.

Petersburg

Institute of Nuclear Physics, Russian Academy of Sciences, St.

Petersburg, Russia.

Structural study of RecA proteins. Structure-function analysis of the

RecA-like protein family in bacteria.

1974-1987 Research Assistant and later Staff Scientist in the

Department of Biophysics at St. Petersburg Technical University,

St. Petersburg, Russia

the role of the rec F gene in homologous recombination. Study of the

characterization of native supercoiling of DNA in bacteria. Generation

of deletion mutants of Topoisomerase I gene.

Experimental Experience

. Isolation and purification of DNA and RNA, DNA cloning and sequencing,

SNPs

. Cloning and overexpression of genes in E.coli, yeast and mammalian cells

. Construction and optimization of protein expression vectors

. Construction and screening of cDNA and genomic DNA libraries (plasmid,

cosmid, bacteriophage, and phage display libraries)

. Site-directed and total (shuffling) mutagenesis

. PCR and RT-PCR techniques, QPCR, Nonradioactive and radioactive methods

of nucleic acids labeling, Hybridization of nucleic acids: Northern and

Southern blot

. Protein purification and analysis, affinity-tag modifications, protein

engineering, protein modeling, protein-protein interactions, 2D gel

electrophoresis

. Various chromatography techniques, including FPLC and HPLC

. Quantification of enzymatic activities, signal transduction assays

. Various microbiology techniques (E.coli, bactriophages, yeast), cell

cultivation, optimization of the target protein expression

. Immunology techniques: FACS, immunoprecipitation, epitope mapping,

Western blot analysis, ELISA, Phage display, antibody design

. Cell culture methods; Stable line generation; Primary cell culture; DNA

transfection, nucleofection, optimization of the target protein expression

. Sindbis and lentivirus RNA/DNA delivery systems, Ad and AAV,

retroviruses, dengue, BSL-2.

. Confocal microscopy, FRET, BRET, fluorescent protein-tag modifications.

. Skillful in using various computer programs

Publications and presented papers:

3. Mirkin, S.M.; Zaitsev, E.N.; Paniutin, I.G.; Lyamichev, V.I. (1984)

Native supercoiling of DNA: The effect of DNA gyrase and omega protein in

E.coli. Mol.Gen.Gent. 196; 508-512.

8. Bachlanova, I.V.; Alexeev, A.A.; Zaitsev E.N.; Zaitseva E.M;

Stepanova, I.M.; Lanzov V.A. (1991)

Functional characteristics of the recA gene from Serratia marcescens Sb.

Gene 101(1):139-141

9. Zaitsev, E.; Alexseyev, A.; Lanzov, V.; Satin, L.; Clark A.J. (1994)

Nucleotide sequence between recA and alaS in E.coli K12 and the sequence

change in four recA mutations. Mutation Research 323:173-177.

11. Alexsejev, A.A.; Bakhlanova,I.V.; Zaitsev,E.N.; Lanzov,V.A. (1996)

Genetic characteristics of new recA mutants of Escherichia coli K-12. J

Bacteriol. 178: 2018-2024.

12. Zaitsev EN, Kowalczykowski SC. (1998) Binding of double-stranded DNA

by Escherichia coli RecA protein monitored by a fluorescent dye

displacement assay. Nucleic Acids Res. Jan 15;26(2):650-4.

13. Zaitsev EN, Kowalczykowski SC. (1998) Essential monomer-monomer

contacts define the minimal length for the N-terminus of RecA protein.

Mol Microbiol. Sep;29(5):1317-8.

14. Zaitseva EM, Zaitsev EN, Kowalczykowski SC. (1999) The DNA binding

properties of Saccharomyces cerevisiae Rad51 protein. J Biol Chem. Jan

29;274(5):2907-15.

15. Zaitsev EN, Kowalczykowski SC. (1999) The simultaneous binding of two

double-stranded DNA molecules by Escherichia coli RecA protein. J Mol

Biol. Mar 19;287(1):21-31.

16. Zaitsev EN, Kowalczykowski SC. (1999) Enhanced monomer-monomer

interactions can suppress the recombination deficiency of the recA142

allele. Mol Microbiol. Oct;34(1):1-9.

17. Zaitsev EN, Kowalczykowski SC. (2000) A novel pairing process

promoted by Escherichia coli RecA protein: inverse DNA and RNA strand

exchange. Genes Dev. Mar 15;14(6):740-9.

18. M. F. Egan, M. Kojima, J. H. Callicott, T. E. Goldberg, B. S.

Kolachana, E. Zaistev, A. Bertolino, B. Gold, D. Goldman, M. Dean, B. Lu,

and D. R. Weinberger. (2003) The BDNF val66met polymorphism affects

activity-dependent secretion of BDNF and human memory and hippocampal

function. Cell 112, 257-269. With Preview by John Garbriele and Alison

Preston, Cell 112, 144-145

19. Dmitry M. Baitin, Eugene N. Zaitsev, and Vladislav A. Lanzov. (2003)

Hyper-recombinogenic RecA protein from Pseudomonas aeruginosa with

enhanced activity of its primary DNA binding site.

J Mol Biol. Apr 18;328(1):1-7.

20. Du J., Feng L., Zaitsev E., Lui X., and Lu B. (2003) Activity-

dependent regulation of TrkB Receptor Tyrosine Kinase and its

internalization in hippocampal neurons. J Cell Biol. Oct 27;163(2):385-

95.

21. P. T. Pang, H. K. Teng, N. Woo, E. Zaitsev, K. Sakata, S. Zhen, K. K.

Teng, W.-H. Yung, B. L. Hempstead, and B. Lu. (2004) Cleavage of ProBDNF

by the tPA/plasmin is essential for long-term hippocampal plasticity.

Science. 306, 487-491.

22. H. Lou, S.K. Kim, E. Zaitsev, C.R. Sneil, B. Lu, and Y.P. Loh. (2005)

Sorting and activity dependent secretion of BDNF requires an interaction

with the sorting receptor Carboxypeptidase E. Neuron 45, 245-255.

23. G. Nagappan, E. Zaitsev, V. V. Senatorov, J. Yangb, B. Hempstead, and

B. Lu. (2009) Control of extracellular cleavage of ProBDNF by high

frequency neuronal activity. PNAS 106, 1267-127.

24. Koshimizu H, Kiyosue K, Hara T, Hazama S, Suzuki S, Uegaki K,

Nagappan G, Zaitsev E, Hirokawa T, Tatsu Y, Ogura A, Lu B, Kojima M.

(2009) Multiple functions of precursor BDNF to CNS neurons: negative

regulation of neurite growth, spine formation and cell survival. Mol

Brain. 13;2(1):27

25. Eugene Zaitsev, Guhan Nagappan, Joseph Callicott, Andreas Meyer-

Lindenberg, Feng Yang, Hyun-Soo Je,

Daniel Weinberger, and Bai Lu (2010) Polymorphismin 5'UTR of Human

BDNF Gene Alters Cellular Distribution of BDNF and Hippocampal

Function. Manuscript in preparation.

26. B. Lu, G. Nagappan, E. Zaitsev. US Patent; U.S. provisional Application

S/N: 60/816,597. Title: Method For Measuring Mature BRAIN DERIVED

NEUROTROPHIC FACTOR.

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