Curriculum
vitae
Elena
María Hilario Andrade
Address Landcare
Research, Private Bag 92-170, 120 Mt
Albert Rd. Mt Albert, Auckland New
Zealand
Telephone 64-9-815-4200
x7024
FAX 64-9-849-7093
e-mail hilarioe@landcare.cri.nz
Personal Born
April 30, 1966, México City, México.
Education
Ph D degree 1/1998 University
of Connecticut, USA (Cell Biology).
Title of Dissertation: The vacuolar H+-ATPase from Giardia lamblia: a potential model
for the study of the evolution of the first eukaryotes. Advisor: Prof. Dr. J. Peter Gogarten.
Masters Degree 12/ 1992 UNAM
(Universidad Nacional Autónoma de México) (Biochemistry). Title of Dissertation: Characterization of
the cytoplasmic and membrane bound pyrophosphatases from the cyanobacterium Spirulina
maxima. Advisor: Prof. Dr.
Heliodoro Celis Sandoval.
Diploma 1/1990 UNAM
(Biology). Title of Dissertation:
Determintation of the physicochemical constants and spectrophotometric
characteristics of the metallochormic Zincon and its potential use in
biological systems. Advisror: Prof. Dr.
Heliodoro Celis Sandoval.
Employment
2001-present Scientist, Plant gene mapping
1998-2001 Postdoctoral fellow at Landcare
Research, New Zealand
1997 Teaching Assistant in
Advanced Biochemistry Laboratory, Dept. Molecular Cell Biology, University of
Connecticut, USA.
5/1990-1/1993 Teaching assistant in Biochemistry
course (laboratory), UNAM
Professional
activities
Member of the New Zealand Microbiological Society
Member of the American Society for Microbiology
Member of the Mexican Society of Biochemistry
Member of the International Society for the Study of
the Origin of Life (ISSOL) and New Zealand representative of ISSOL.
Committee member in oral defenses (diploma in Biology,
UNAM) 1991-1993
Fellowships
10/1999 Travel
grant to attend the Gordon Research Conference on Molecular Evolution, Hayama,
Japan.
6-7/1995 Recipient
of an Award from the Dept. of Navy, Office of Naval Research to attend the
Microbial Diversity Course at the Marine Biological Laboratory, Woods Hole, MA.
6-7/1994 Recipient
of the NASA Planetary Biology Internship
7/1993 Recipient
of a travel grant from ISSOL.
*1/1993-1/1997 Ph
D. studies at University of Connecticut, USA.
*11/1990-12/1992 Master
degree studies at UNAM, México.
*11/1989-10/1990 Prerequisite
courses for master degree studies, UNAM.
*10/1988-9/1989 Undergraduate,
Biology at UNAM.
*1983-1985 Private
highschool, México.
*
from UNAM
Publications
HilarioE.
Labelling and Detection, in "Gene Probes -Principles and Protocols"
Methods in Molecular Biology Series. (Aquino de Muro M. and Rapley R. eds),
Humana Press, Totowa, NY. (in preparation)
Hilario E,
Gogarten JP. (1998) The prokaryote to eukaryote transition reflected in the
evolution of the V/F/A-ATPase catalytic and proteolipid subunits. J. Mol. Evol.
46:703-715
Olendzenski L, Hilario E, Gogarten JP. (1998).
Horizontal Gene Transfer and Fusing Lines of Descent: The Archaebacteria -a
Chimera? in "Horizontal Gene
Transfer", M. Syvanen and C. Kado (eds.), Chapman/Hall, London, pp 349-362
Gogarten JP, Olendzenski L, Hilario E. (1998)
Gene Duplications and Horizontal Gene Transfer. In : "Thermophiles: the
key to molecular evolution and the origin of life?" eds. Michael Adams and
Juergen Weigel, publ.; Taylor and Francis, pp. 165-176
Gogarten JP, Hilario E, Olendzenski L. (1997)
The Tree of Life. American Society for Microbiol. News 63, 404-405
Gogarten JP, Olendzenski L, Hilario E, Simon C,
Holsinger KE. (1996). Dating the Cenancester of Organisms - Technical Comment.
Science 274 (5293):1750-1751
Gogarten JP, Hilario E, Olendzenski L (1996).
Gene duplications and Horizontal Gene Transfer during Early Evolution, In
"Evolution of Microbial Life" (Roberts McL., Sharp P, Alderson G,
Collins M, eds.) Soc. Gen. Microbiol. Symp.
54. Cambridge Press
Hilario E, Gogarten JP (1995) The V-ATPase A subunit
gene (vma-1) from Giardia lamblia. Biochem. Biophys. Acta 128,
94-98
Gogarten-Boekels M, Hilario E, Gogarten JP.
(1995) The effects of heavy meteorite bombardment on the early evolution- The
emergence on the three domains of life. Origins of Life and Evolution of the
Biosphere. 25, 251-264
Hilario E,
Gogarten JP (1993) Horizontal transfer of ATPase genes -the tree of life
becomes a net of life. Biosystems 31,
111-119
Hilario E, Romero
I, Celis H. (1990) Determintation of the physicochemical constants and
spectrophotometric characteristics of the metallochormic Zincon and its
potential use in biological systems. J. Biochem. Biophys. Meth. 21,
197-207
Scientific
Conferences
Hilario
E and Gogarten JP. The Intein in the Thermoplasma
A-ATPase A subunit: Evolutionary and functional implications. IX International Congress of Bacteriology
and Applied Microbiology, Sydney, Australia. August, 1999. And Gordon Conference on Molecular Evolution,
Hayama, Japan, October, 1999.
Hilario
E and Young JM. Gene Transfer among Pseudomonas
spp. from saprophytic plant associations.
IX International Congress of Bacteriology and Applied Microbiology,
Sydney, Australia. August, 1999. And
Gordon Conference on Molecular Evolution, Hayama, Japan, October, 1999.
Hilario
E and Gogarten JP. The prokaryote to
eukaryote transition reflected in the evolution of the V/F/A-ATPase catalytic
and proteolipid subunits. Gordon
Research Conference on Microbial Population Biology, Plymouth, New
Hampshire, July 27-August 2, 1997. Also
presented as contributing speaker at the New England Molecular and
Evolutionary Biology Meeting. Yale Universtity, November 1997, and at the NZMS/NZSBMB
Microbes and Molecules Meeting, Masterton, New Zealand, Nov 30-Dec 3, 1998.
Hilario
E and Gogarten JP. The vacuolar
H+-ATPase from Giardia lamblia: a potential model for the study of the
evolution of the first eukaryotes. Presented at the New England Molecular
and Evolutionary Biology Meeting. November 1995, Storrs CT. Also presented at the Eastern Great Lakes
Molecular Evolution Meeting, May 1996, Cornell University, NY.
Hilario
E and Gogarten JP. Horizontal
transfer of ATPase genes -the tree of life becomes a net of life. Presented at
the New England Molecular and Evolutionary Biology Meeting. November
1993, New York, N.Y., and the Gordon Research Conferences in Origin of Life,
New Port, RI July 1994.
Hilario,E, Gogarten JP, Sosa-Peinado A, Silva E, Lazcano A.
Early Gene Duplication and the Origin of Biochemical Regulation. 10th
International Conference on the Origin of Life. Barcelona, Spain July 1993.
Hilario
E, Gómez Lojero C, Celis H. The
membrane-bound pyrophosphatases of the cyanobacterium Spirulina maxima. VII
Panamerican Asocciation of Biochemistry Societies Conference. Ixtapa, Gro.
México September 27-October 2, 1992.
Celis H,
Hilario E, Romero I. Caracterización del metalocrómico Zincon (óxido
2-carboxi-2'-hidroxi-5-sulfoformazilbenceno) y su utilización en sistemas
biológicos. I Iberoamerican Congress on Biophysics. Sevilla, Spain,
September, 1989.
Research
Experience
Phylogenetic analysis of nucleotide and amino acid
sequences using the following programs: PCGene, ClustalW, Sequencher, Phylip,
Paup*, Puzzle, Statalign, as well as accessing data bases through the World
Wide Web.
Molecular biology techniques; genomic DNA extractions
from bacteria and Giardia; restriction pattern analysis; Southern blots
and library screening using non-radioactive probes and colorimetric detection
methods; primer design and PCR techniques; DNA sequencing and editing; genomic
subtraction protocols.
Basic techniques on protein purification: crude
cytoplasmic and membrane preparations; size exclusion, ionic exchance and
affinity chromatographies; native and denaturing electrophoresis
Bacterial cultures of acidothermophiles, halophiles,
mesophiles and photosynthetic bacteria.
Axenic cultures of the protist Giardia lamblia
Antibody production in chickens, extraction of IgY and
affinity purification of specific antibodies; Western blot analysis
Enzymatic activity determination of pyrophosphatases
and H+-ATPases
Single and double beam spectrophotometry
Current
Research Project
My main research programme is the impact of horizontal
gene transfer in microbial evolution.
My postdoctoral research aims to search for evidence of horizontal gene
transfer among saprophytic Pseudomonas species. This study includes twenty-two Pseudomonas
strains and the project includes two approaches. The first one consists of obtaining partial sequences of three
molecular markers: recombinant protein A (recA), the small subunit of
carbamoyl phosphate synthase (carA), and the catalytic subunit of the
F0F1-ATP synthase (atpD). Based
on the codon usage and G+C content of the genus Pseudomonas, I designed
redundant oligonucleotides to amplify and sequence the core region of each
marker. I am currently carrying out the
phylogenetic analysis of the three molecular markers of the selected species
plus representatives of the proteobacteria group reported in databases.
The
second approach of this project is an experimental search for lateral transfer
of DNA. I set up the genomic
subtraction technique and constructed a subtraction library for P. agarici
subtracted with ten Pseudomonas strains. Two unique subtraction clones were sequenced. Both contain a significant lower G+C content
(0.45 and 0.52, respectively, compared to 0.61 in coding regions or 0.58-0.7 in
non-coding regions). Southern blot
analysis confirmed that both clones belong to the subtracted species and are
absent in the other ten subtracter species.
The clone with the lowest GC content was used as probe to isolate a
larger restriction fragment for further characterization. A 3.1 kb fragment was cloned and partially sequenced. An open reading frame similar to the
macrophage infectivity potentiator protein of several proteobacteria has been
identified. Charaterization of the
upstream and downstream regions is now under study.
Another
research interest related to horizontal gene transfer is the importance of
mobile genetic elements in the establishment and maintenance of microbial
communities. In collaboration with Dr
Jacqueline Aislabie (Landcare Research New Zealand) we are preparing a research
programme proposal to apply molecular techniques such as terminal restriction
fragment length polymorphism (T-RFLP) based on 16S-rRNA and other functional
genes. We will compare the genetic
diversity of microbial communities of Antarctic soils from three locations at a
range of depths, and monitor changes in the microbial community of soil
following exposure to hydrocarbons. I
have been invited to participate in a grant proposal for the NASA Astrobiology
Institute, lead by my PhD advisor Dr Peter Gogarten. This proposal will study horizontal gene transfer as a historical
process, as well as an experimental approach to evaluate the dynamics of mobile
genetic elements in microbial populations.
A
fortunate outcome of my PhD research thesis was the discovery of a small intein
located in the catalytic subunit of the A-ATP synthase of Thermoplasma acidophilum. I have followed up this study with Dr Peter
Gogarten, and presented an updated compilation of small inteins last year at
the IX International Congress of Bacteriology and Applied Microbiology, Sydney,
Australia.
I enjoy
the full process of scientific research: from proposing new ideas and
hypothesis, obtaining the data, troubleshooting daily problems in the
experimental designs, up to the analysis and discussion of the results. My laboratory skills have undoubtedly helped
me to appreciate the complete picture of science.
References
1. Prof
Dr Johann Peter Gogarten
Department
of Molecular and Cell Biology
University
of Connecticut
75
N. Eagleville Rd. U-44
Storrs,
CT 06269-3044 USA
Phone 1-860-486-4061
Fax(860) 1-860-486-1784
Email gogarten@uconnvm.uconn.edu
2. Dr
Richard D. Newcomb
Science
Leader, Plant Health and Development Group
The
Horticulture and Food Research Institute of New Zealand Ltd.
Mt
Albert Research Centre, 120 Mt Albert Road,
Private
Bag 92-169 Auckland, New Zealand
Phone 64-9-8154200 Ext 7294
Fax 64-9-8154227
email rnewcomb@hort.cri.nz
3. Dr
Ross E. Beever
Former
Programme Leader
Landcare
Research, Ltd.
Mt
Albert Research Centre, 120 Mt Albert Road,
Private
Bag 92-170 Auckland, New Zealand
Phone 64-9-8154200 Ext 7310
Fax 64-9-849-7093
email beeverR@landcare.cri.nz