POSTER PRESENTATIONS
P1 Functional Genomics, Proteomics and Bioinformatics
P1–1
The influence of the HERV-K LTR on KIAA1245
subfamily gene expression
N. Abrarova, E. Stoukacheva, T. Vinogradova and E. Sverdlov
Laboratory of structure and function of human genes, Institute of
Bioorganic Chemistry, Moscow, RUSSIA
Long terminal repeats (LTR) of endogenous retroviruses com-
prise about 8% of human genome. Typical LTR contains a set
of regulatory elements: promoters, enhancers, polyadenilation
sites, which can take part in neighbouring genes expression regu-
lation. Earlier, we have described a subfamily of closely related
genes highly similar to the KIAA1245 mRNA, part of which
contains a HERV-K LTR in their structure, whereas the other
lacks it. Using this subfamily as a model for studying regulation
of gene transcription, we showed that LTR serves as an alterna-
tive promoter and possess high enhancer activity. Genes that
contain LTR differ from genes lacking it in cell type specificity of
transcription. We generated a series of successive 5¢- and 3¢-dele-
tion mutants with a 200 bp increment, and also two middle vari-
ants, 200 and 400 bp in length (Mid200 and Mid400,
respectively). To determine promoter activity of LTR and its
fragments they were cloned into pGL3-BV vector, containing
luciferase reporter gene. The full-sized LTR demonstrated high
promoter activity in Tera1 and NT2/D1 cell lines. To determine
the enhancer activity LTR and subfragments were cloned in
pGL3-PV vector, which has SV40 promoter in its structure
besides luciferase gene. Transient transfections demonstrated that
Mid200 fragment of the LTR exhibits high cell type specific
enhancer activity. In Tera1 cell line it was comparable to the
activity of universal SV40 enhancer. This fact allows to suggest
that enhancer is localized in this region of the LTR. The analysis
of transcription of KIAA1245 subfamily genes have shown that
LTR-lacking gene Al592309 is not transcribed in Tera1 and
NT2/D1 cell lines, whereas LTR-containing genes are transcribed
in these cell lines. Thus, we may suggest that insertion of the
LTR in second intron of KIAA1245 genes may lead to change in
cell type specificity of their transcription. This fact allows to sug-
gest the role of the LTR in evolution of KIAA1245 subfamily
genes.
P1–2
Study of DNA interactions with cerium (III),
lanthanum (III) and gadolinium (III) ions by
using of Raman spectroscopy
V. Kohoutkova
1
, P. Babula
1
, R. Opatrilova
1
, O. Vrana
2
,
V. Adam
3
, J. Zehnalek
3
and R. Kizek
3
1
Department of Natural Drugs, University of Veterinary and
Pharmaceutical Sciences, Brno, CZECH REPUBLIC,
2
Institute of
Biophysics, Academy of Sciences of the Czech Republic, Brno,
CZECH REPUBLIC,
3
Department of Chemistry and Biochemis-
try, Mendel University of Agriculture and Forestry, Brno, CZECH
REPUBLIC
The biological properties of the lanthanides, based on their simi-
larity to calcium, have stimulated research into their therapeutic
application. Up-to-date we have been successfully using at least
two pharmaceuticals cerium nitrate as a topical cream with silver
sulfadiazene for the treatment of burn wounds and lanthanum
carbonate as a phosphate binder for the treatment of hyperphos-
phatemia. Lanthanides3+ compounds have also been investi-
gated for their anti-cancer potential. Clinical reports suggested
that several compounds such as cerium (III) iodide posed cyto-
toxic effect, however the biochemical mechanism of their action
is still unclear (1–3). Therefore, we aim our attention on study of
interactions DNA with inorganic salts of cerium (III), lanthanum
(III) and gadolinium (III). Raman spectroscopy was employed to
characterize the perturbations to DNA conformation induced in
DNA by three inorganic salts of the above mentioned lantha-
nides (2). All studied lanthanides coordinated to N7 of two
neighbouring guanine bases, as this nitrogen does not form H
bonds with other bases, in the same or in opposite DNA strands.
The results of the present work demonstrate that Raman spec-
troscopy represents a suitable tool to provide insights into struc-
tural factors involved in the mechanisms underlying antitumor
effects of drugs.
Acknowledgement: This work was supported by GA CR 522/
07/0692.
References:
1. Fricker SP. Chem. Soc. Rev. 2006; 35(6): 524–533.
2. Vrana O et al.,J. Struct. Biol. 2007; 159(1): 1–8.
3. Babula P et al.,Curr. Pharm. Anal. 2009; 5(1): 47–68.
P1–3
Real-time polymerase chain reaction with
electrochemical detection for detection
pandemic viruses
V. Adam
1
, D. Huska
1
, S. Krizkova
1
, J. Hubalek
2
and R. Kizek
1
1
Department of Chemistry and Biochemistry, Mendel University of
Agriculture and Forestry in Brno, Brno, CZECH
REPUBLIC,
2
Department of Microelectronics, Brno University of
Technology, Brno, CZECH REPUBLIC
Viral infections pose a threat for man kind. Diagnostic systems
focus on the direct cultural proof of identity. Almost all systems
use polymerase chain reaction. Real-time polymerase chain reac-
tion is based on the polymerase chain reaction and routinely used
to amplify and simultaneously quantify a targeted DNA molecule.
Fluorescent dyes intercalating with double-stranded DNA and
modified DNA oligonucleotide probes fluorescing when hybridized
with a complementary DNA are used for real time quantification
of the amplicons. The process of DNA quantification has demands
on robust instrument and on high cost chemicals. Electrochemical
detection is low cost and sensitive alternative method to detect
nucleic acids (1, 2). The main aim of this work is to propose novel
electrochemical detector for monitoring PCR. Miniaturized poten-
tiostat with nanotubes-based screen-printed electrodes was used
for detection of specific sequence of severe acute respiratory syn-
drome, avian influenza virus and Ebola virus amplified by poly-
merase chain reaction. The amplicons were obtaining after 2, 4, 6,
8, 10, 15, 20, 25, 30 and 35 cycles, whereas the amount of amplified
DNA was between 10 and 100 pg. The amplicons obtained even
after two cycles were detectable by using the electrochemical
instrument. Moreover we compared the results with agarose gel
electrophoresis. The lowest detectable amount of DNA was
obtained after 15 PCR cycles.
Acknowledgement: This work was supported by GA AV
KAN208130801.
Poster Presentations Abstracts
FEBS Journal 276 (Suppl. 1) 95–356 (2009) ª2009 The Authors Journal compilation ª2009 Federation of European Biochemical Societies 95
References:
1. Palecek E et al. Anal. Chim. Acta. 2002; 496(1): 73–83.
2. Huska D et al. Electrophoresis 2008; 29(24): 4964–4971.
P1–4
Spatial distribution of heavy metals in healthy
and melanoma animal tissues
V. Adam
1
, O. Zitka
1
, D. Huska
1
, S. Krizkova
1
, J. Strnadel
2
,
V. Horak
2
, T. Vaculovic
3
, K. Novotny
3
, V. Kanicky
3
, J. Kaiser
4
and R. Kizek
1
1
Department of Chemistry and Biochemistry, Mendel University of
Agriculture and Forestry in Brno, Brno, CZECH
REPUBLIC,
2
Academy of Sciences of the Czech Republic, Insti-
tute of Animal Physiology and Genetics, Libechov, CZECH
REPUBLIC,
3
Department of Chemistry, Masaryk University,
Brno, CZECH REPUBLIC,
4
Institute of Physical Engineering,
Brno University of Technology, Brno, CZECH REPUBLIC
Levels of some of heavy metals are monitored and their altera-
tions can be used as diagnostic markers. Nevertheless, spatial dis-
tribution of the essential metals in animal tissues is still not clear.
The aim of this work is detection of copper and zinc in healthy
and tumour tissues of miniature pigs by using of the laser
induced breakdown spectroscopy (LIBS). Concentration of essen-
tial-heavy-metal-transporting protein metallothionein (MT) was
determined by the Brdicka reaction. Tissue cryosections (thick-
ness 40 lm) were obtained from the MeLiM strain of miniature
pigs with hereditary melanoma, particularly from healthy skin,
cutaneous nodular melanomas and metastases in the liver, spleen
and lymph nodes. Using LIBS we measured maps of spatial dis-
tribution of the essential heavy metals in cryosections and found
that the maps of healthy and tumour cryosection markedly dif-
fered. The highest content of MT was determined in the tumours
localised on the back of animals and was nearly 500 lgofMT
per gram of tissue. The MT levels determined in metastases in
spleen, lungs and liver were within the range from 40 to 160 lg/g
of the tissue, the average level was 110 ± 40 lg/g of the tissue.
Acknowledgements: This work was supported by the GA AS
CR (IAA401990701) and by the Ministry of Education, Youth
and Sport CR (2B08063) and Liga proti rakovine Praha (2009).
References:
1. Krizkova S et al. Sensors 2008; 8(5): 3106–3122.
2. Kaiser J et al. Spectrochim. Acta Part B 2009; 64(1): 67–73.
P1–5
Chromatin accessibility of MHCII locus and
enhanceosome stability determine
transcriptional competence through mitosis
P. Arampatzi, M. Gialitakis, T. Makatounakis and
J. Papamatheakis
Department of Biology, Institute of Molecular Biology and
Biotechnology, Heraklion, GREECE
During mitosis, transcription factors and RNA polymerase II are
displaced from mitotic chromatin, which is condensed and tran-
scription is interrupted. However specific gene regions are sug-
gested to be bookmarked by TBP and histone modifications for
rapid reactivation after cell division. The MHCII locus is trans-
criptionally activated by the assembly of an enhanceosome
(MCE) which recruits the master regulator, CIITA. We found
that endogenous or GFP-fusions subunits of the MCE remain
associated and dynamically exchanged on mitotic chromosomes.
Chromatin immunoprecipitation assays show significant occu-
pancy by the MCE, CIITA and general transcriptional machinery
at various phases of the cell cycle including mitosis in B lym-
phoid cells. Mitotic maintenance of the MCE does not depend
on CIITA expression and correlates with a chromatin state that
is fully or partly maintained in mitotic lymphoid or non lym-
phoid cells, respectively. Monitoring of newly synthesized RNA
shows reduced transcriptional activity of the DRagene in mito-
sis. Constitutive expression of exogenous CIITA is sufficient to
fully support mitotic expression in lymphoid but not in epithelial
cells. Using an inducible system we show that CIITA synthesised
in cells arrested in mitosis rescues MHCII transcription to asyn-
chronous levels in lymphoid cells demonstrating full expression
competence of DRagene. These results show there are two pat-
terns of mitotic deficiency of gene expression: in B lymphoid cells
low but significant MHCII transcription can be fully rescued, as
opposed to epithelial cells that show minimal mitotic transcrip-
tion may due to enhanced chromatin condensation.
P1–6
Associations between PTGS1 gene mutations
and aspirin resistance in patients with
congenital heart diseases
I. Coskun
1
, Y. Atay
1
, A. Berdeli
2
, M. Mecidov
1
, A. Atay
3
,
M. F. Ayik
1
, T. Yagdi
1
and E. A. Alayunt
1
1
Cardiovascular surgery, Ege University, izmir, TURKEY,
2
Pediatric Molecular Medicine Laboratory, Ege University, izmir,
TURKEY,
3
Department of Clinical Biochemistry, Ataturk Training
Hospital, izmir, TURKEY
Introduction: This study was aimed to evaluate that frequency
and existance of PTGS1 gene mutation which coded cyclooxy-
genase-1 (COX-1) enzyme and associations with aspirin resistance
in patients with congenital heart diseases (CHD) in Turkish pop-
ulation.
Methods: It was included 90 patients with CHD treated by
aspirin after their operations. 24 healthy participants were
enrolled as control group. Direct DNA coding sequence method
was used for analysing PTGS1 gene mutation. PCR amplification
was made by using specific oligo primers (MWG) and AmpliTaq
Gold PCR kits for four exons. After capillary gel electrophoresis,
nucleotide sequences of PTGS1 gene was obtained and compared
with cDNA Genbank (NM_000962) and protein (NP_000953)
sequences to determine single- nucleotide polymorphisms and
amino acid mutations.
Results: W8R missense aminoacid mutation was the most fre-
quent mutation (83.3%) in exon-2. P188P sinonym aminoacid
polimorphism (34%) and D189E amino acid mutation (20%)
were other mutations detected in exon-6 in patient group. In con-
trol group, W8R missense aminoacid mutation (83.3%) in exon-
2, Q41Q sinonim aminoacid mutation (8.3%) in exon-3, D189E
missense aminoacid mutation (12.5%) in exon-6 were found.
Nine new missence amino acid mutations (L13P; L15V; P18R;
C58T; I45M; R53S; D189E; F200L; R239L) and eight sinonym
amino acid mutations (L23L; P39P; R53R; G44G; G62G;
K185K; P18P) were detected. It was observed the association
between aspirin resistance and missence amino acid mutations in
patients.
Conclusion: Genetic variations in PTGS-1 may affect aspirin
resistance in CHD. Detecting these variations may help to predict
drug responce and to select optimal therapy and dosage regi-
mens. This is the first study in our country dealing with PTGS1
gene mutation and polymorphism which coded COX-1 enzyme
and association with aspirin resistance in CHD.
Abstracts Poster Presentations
96 FEBS Journal 276 (Suppl. 1) 95–356 (2009) ª2009 The Authors Journal compilation ª2009 Federation of European Biochemical Societies
P1–7
Analysis of the ID1 gene expression level and
promoter methylation in patients with benign
and malignant thyroid nodules
A. Banaszewska
1
, M. Piechota
1
, K. Drobnik
2
, K. Ziemnicka
2
,
K. Ziemnicki
1
and R. Plewa
1
1
Department of Animal Physiology and Development, Adam
Mickiewicz University, Poznan, POLAND,
2
Department of
Endocrinology Metabolism and Internal Disease, University of
Medical Science, Poznan, POLAND
Inhibitor of DNA binding (ID1) has been proposed to serve a
dual function as a regulator of cell proliferation and differentia-
tion in both normal and pathological conditions associated with
cancer development. ID is a transcription family of factors lack-
ing basic DNA binding domain, which form inactive heterodi-
mers with basic helix-loop-helix transcription factors and inhibits
expression of tissue specific genes and cells differentiation. Recent
research shows, that ID1 mRNA level correlates with malignant
thyroid carcinoma phenotype. Thus, dual role of ID1 gene in
benign and malignant thyroid nodules was studied. ID1 mRNA
level of 33 patients was investigated in human papillary thyroid
carcinoma (PTC), non-toxic nodular goiter, toxic nodular goiter
and nodular goiter with Hashimoto disease using Real time PCR
and Delta Delta Ct method to estimate relative amount of ID1
mRNA. Furthermore, genomic DNA was extracted and ID1 pro-
moter methylation was examined in the majority of patients spec-
imens. Six of seven PTC patients samples were found to have
barely detectable ID1 mRNA level. Expression of ID1 was higher
in non-toxic nodular goiter and in nodular goiter with Hashimoto
disease in comparison to PTC and toxic nodular goiter
(P< 0.05). No hypermethyleted ID1 gene promoter was found
in the majority of analyzed cases. In our study no significant
association was observed between ID1 gene expression and
malignant thyroid nodules. This findings indicate that ID1 could
not be a marker of aggressive phenotype in PTC.
P1–8
TNF alpha -308 polymorphism and matrix
metalloproteinase-9 level in children with
juvenile idiopathic arthritis threated with
etanercept
J. Basic
1
, D. Pavlovic
1
, T. Jevtovic-Stoimenov
1
, J. Vojinovic
2
,
M. Marinkovic
1
and A. Veljkovic
1
1
Medical Faculty, Institute of Biochemistry, Nis, SERBIA,
2
Clinic
of Pediatrics, Clinical Centre Nis, Nis, SERBIA
Juvenile idiopathic arthritis (JIA) is the most common form of
persistent arthritis in children. Tumor necrosis factor-a(TNF- a)
is likely to have a primary role in the pathogenesis of JIA,
including matrix metalloproteinase-9 (MMP-9) production. The
aim of this study was to investigate G-A -308 single nucleotide
polymorphism (SNP) in the promoter region of TNF agene, as
well as to evaluate the eventual influence of etanercept (TNF
receptor II-Fc fusion protein) on MMP-9 level in children with
JIA. A total of 42 patients with JIA were screened for the poly-
morphism using the PCR-RFLP method. Plasma MMP-9 level
was determined before starting etanercept therapy and 12 months
after, using Elisa kit. The genotype -308GG was present in 13
(31%) patients and the -308AA genotype was present in two
(4.7%) patients. Twenty-seven (64.3%) patients were heterozy-
gous. MMP-9 level in patients with the genotype -308GG was
significantly decreased after 1 year of treatment with etanercept
compared to the value before (P = 0.01). On the other hand,
there were no significant decrease of MMP-9 levels after treat-
ment in patients with the genotype -308GA and -308 AA
(P = 0.147; P = 0.126). We discussed decrease of MMP-9 level
in children with -308GG genotype as a possible consequence of
better response to etanercept treatment compared to A allele car-
riers.
P1–9
Screening of Three Exons of the RET Proto-
oncogene in Turkish Patients with Papillary
Thyroid Carcinoma
N. S. Bayramci
1
, L. Acik
2
, L. Y. Koc
3
, G. Vural
4
, M. Kilic
5
,
M. Tez
5
, M. Koc
5
and N. Ercakmak
4
1
Faculty of Education, Department of Science Education, Bayburt
University, Bayburt, TURKEY,
2
Faculty of Arts and Science,
Department of Biology, Gazi University, Ankara, TURKEY,
3
Faculty of Science, Department of Biology, Ankara University,
Ankara, TURKEY,
4
Nuclear Medicine Department, Dr. Abdurrah-
man Yurtaslan Ankara Oncology Education and Research Hospi-
tal, Ankara, TURKEY,
5
5th Surgery, Ankara Numune Education
and Research Hospital, Ankara, TURKEY
Different types of tumors can develop in thyroid gland cells. Thy-
roid carcinoma represents the most frequent form of cancer of
the thyroid glands, with prevalence in Europe of about 3 per
100 000. PTC is the most frequent histotype of differentiated thy-
roid cancer. The RET proto-oncogene, localized to chromosome
subband 10q11.2, comprises 21 exons, which encodes the protein
RET, a receptor tyrosine kinase (RTK) expressed in derivatives
and tumors of neural crest origin. The RET proto-oncogene is
involved in the genesis of papillary thyroid carcinoma. To deter-
mine the mutation frequency of RET proto-oncogene in Turkish
papillary thyroid carcinoma patients, we conducted this retro-
spective study. Eighty-two patients with PTC were screened for
mutations in exon 10, 11 and 13 using PCR and sequence analy-
sis. A negative control was included in each amplification analy-
sis. Twelve of the patients exhibited mutation, 24 of them
showed SNP. Twelve different mutations located in exon 11 and
three different mutations in exon 13. 12.19% of the patients had
mutation in exon 11. 3.65% of the patients had mutation in exon
13. Twenty-four of the patients exhibited SNP (29.26%). We
found the SNP at codons 631 (1.21%), 769 (1.21%), 691 (3.65%)
in exon 11 and at codons 769 (24.39%) and 763 (1.21%) in exon
13. The most frequent SNP that is found in patients with PTC
(24.3%) involve codon 769 (exon 13) in the tyrosine kinase
domain. The common mutation found in patients involves codon
630 (four patients) in the extracellular domain of RET encoded
by exon 11, less frequently, mutation occurred codons 765, and
770 and 795 (exon 13) in the tyrosine kinase domain. The muta-
tions were found in 41.46% of all thyroid carcinoma patients.
The numbers of mutations were higher in exon 11 than exon 13.
In our series, we detected that four patients in 82 of all PTC
patients presented Cys630Ser (TGC fiAGC, 5.9%) is the most
common mutation at codon 630 in exon 11. In addition to that,
Cys630Ser mutation, is associated with medullary thyroid carci-
noma, however in our genetic testing it was diagnosed on four
patients with papillary thyroid carcinoma. Therefore, the rela-
tionship between Cys630Ser variation and development of PTC
should be further explored. All of the investigated polymor-
phisms are silent mutations, leave out codon 691 polymorphism.
We observed the codon 691 polymorphism in three patients
(GGT/AGT) of all 82 PTC patients. In the 13rd exon region of
the RET proto-oncogene, we found that these possible mutations,
which expressed as an alteration in the form of TCC (serine)
replacement with TGC (sistein) at codon 765, CGA (arginine)
Poster Presentations Abstracts
FEBS Journal 276 (Suppl. 1) 95–356 (2009) ª2009 The Authors Journal compilation ª2009 Federation of European Biochemical Societies 97
replacement with CAA (glutamic acid) at codon 770. The other
mutation in exon 13 was at codon 770, Arg replacement with
Glu. The large number of SNP at codon 769 in exon 13 with
PTC patients was interesting and it should be further explored.
P1–10
Metabolomic profiling of serum from obese
adult females infected by Chlamydophila
pneumoniae
G. Bazylak
1
, C. Ludwig
2
, E. Fagadar-Cosma
3
and
U. L. Gunther
2
1
Department of Pharmaco-Bromatology & Molecular Nutrition,
Faculty of Pharmacy Collegium Medicum Nicolaus Copernicus
University, Bydgoszcz, POLAND,
2
HWB NMR, CR UK Institute
for Cancer Studies, University of Birmingham, Edbagston Birming-
ham, UK,
3
Department of Organic Chemistry, Institute of Chemis-
try Timisoara of Romanian Academy, Timisoara, ROMANIA
Chlamydophila (Chlamydia) pneumoniae is a very common path-
ogen causing an acute respiratory diseases and multidirectional
inflammation progress during a past and persistent infections in
humans of all age. Recently, significant associations between C.
pneumoniane infection and obesity indices have been observed
on results of epidemiological studies in various populations from
several north Europe countries. However, until now the number
of NMR- or MS-based metabolomic and proteomic studies on
specific biomarkers related with variety of nutrition disorders,
including obese subjects, is still scarce. Thus, the goal of our
studies was to find by use of the 1D- and 2D-1H-NMR proce-
dures the range of characteristic, specific and unique low weight
metabolites (proteins) in serum samples from wide group of over-
weight and obese adult females with increased hormonal activity
of adipose tissue, and which indicating past (or persistent) infec-
tion by C. pneumoniae as diagnosed by introductory serological
immunosorbent assay. Proposed here 1H-NMR methodology
should be effective for profiling changes in the serum metabolo-
me (proteome) of subjects with obesity and its associated implica-
tions with bacterial infection and disorders of immunological
status, thus enabling future early diagnosis, risk assessment and
advanced treatment of infection related obesity in humans.
Acknowledgement: Supported by the grant from EU NMR
project – Contract No. RII3-026145.
P1–11
The study on Fnr-type transcription regulators
functions in bacterium Paracoccus
denitrificans using proteomics, transcriptomics
and bioinformatics tools
P. Bouchal
1
, I. Struharova
1
, E. Budinska
2
, O. Sedo
3
,
T. Vyhlidalova
1
, Z. Zdrahal
3
, R. van Spanning
4
and I. Kucera
1
1
Department of Biochemistry, Faculty of Science, Masaryk Univer-
sity, Brno, CZECH REPUBLIC,
2
Institute of Biostatistics and
Analyses, Masaryk University, Brno, CZECH REPUBLIC,
3
Department of Experimental Biology, Masaryk University, Brno,
CZECH REPUBLIC,
4
Department of Molecular Cell Physiology,
Vrije Universiteit Faculty of Earth and Life Sciences, Amsterdam,
THE NETHERLANDS
Paracoccus denitrificans is a facultatively autotrophic soil bacte-
rium which responds to decreasing level of oxygen in the environ-
ment by a switch from aerobic to anaerobic (denitrifying) growth
mode. In our work, we analyzed roles of fnr-type transcription
regulators FnrP, NNR and NarR (with described key roles as
sensors for oxygen, nitrate and nitrous oxide) using proteomics,
transcriptomics and bioinformatics tools to consider their roles in
the adaptation processes at global level. Protein compositions of
four P. denitrificans strains (wild type, FnrP-, NNR- and NarR-
mutants grown aerobically, semiaerobically and semiaerobically
with nitrate) were analyzed using set of 36 large format 2-D
PAGE gels, their statistical evaluation and MS/MS protein iden-
tification of more than 500 proteins. Expression differences of
key up-/down-regulated gene products were validated using qRT-
PCR. Comparative proteomic analysis revealed, besides other
results, differences between strains in the expression of three key
denitrification enzymes (nitrate reductase, nitrite reductase,
nitrous oxide reductase), OmpW and UspA proteins. These find-
ings are in agreement with known mechanisms and/or confirmed
the positions of proposed fnr binding sites in their promoters.
Detection of additional group of up-/down- regulated proteins
without fnr binding sites in their promoters (e.g. SDR dehydro-
genases, TonB receptors) indicates involvement of additional reg-
ulatory mechanisms into the adaptation processes studied.
Evidences for potential roles of transcription regulators of other
families (e.g. LysR, MarR and two-domponent regulators) are
discussed.
Acknowledgements: We are thankful to Czech Science Foun-
dation (grant No. 203/07/P0471) and to Czech Ministry of Edu-
cation (grant No. MSM0021622413) for financial support.
P1–12
Analysis of proteome alteration in tomato
roots after Meloidogyne hapla infection
A. Obrepalska-Steplowska
1
, K. Nowaczyk
1
, M. Luczak
2
,
M. Figlerowicz
2
, R. Dobosz
3
and M. Budziszewska
1
1
Interdepartmental Laboratory of Molecular Biology, Institute of
Plant Protection, Poznan, POLAND,
2
Polish Academy of Sci-
ences, Institute of Bioorganic Chemistry, Poznan, POLAND,
3
Department of Zoology, Institute of Plant Protection, Poznan,
POLAND
Root-knot nematodes from the genus Meloidogyne are parasitic
to many economically important crop plants and cause great
losses in the crop production. Out of fifteen European species,
eight were found in Poland; among them the most common and
widely distributed is M. hapla, parasiting on dicotyledonous
plants. During the initial stage of infection characteristic root
galls form on the roots of susceptible plant hosts. As a result of
interaction between parasite and the host, in the vascular cylinder
of the root permanent feeding sites are formed with specialized
multinucleated feeding cells (giant cells). The redifferentiation of
root cells and development of metabolically active giant cells is
induced by the nematode. The mechanism of giant cells forma-
tion and the role of plant proteins in this processs remain
unclear. Our attempt was to analyse the changes in the plant pro-
teome in the early stage of M. hapla infection. Tomato plants
were grown in isolated conditions and infected with M. hapla.
After isolation of proteins from roots of healthy and infected
plants, as well as from nematodes, two-dimensional electrophore-
sis was performed. The results of experiments were compared to
ensure elimination of nematode proteins from further analysis.
As some qualitative and quantitative changes were observed, cho-
sen proteins were analysed using mass spectrometry. The decrease
of cytosolic and mitochondrial dehydrogenases of cellular respi-
ration and electron transport chain was observed as well as for
proteins related to cytoskeleton (actins), relevant to changes in
the cell’s shape and wound response (annexin). The qualitative
changes are related with proteasome endopeptidase complex,
absent in infected root cells. Nematode proteins secreted during
Abstracts Poster Presentations
98 FEBS Journal 276 (Suppl. 1) 95–356 (2009) ª2009 The Authors Journal compilation ª2009 Federation of European Biochemical Societies
infection had been the subject of many recent investigations. But,
as the relationship between these parasites and the host is very
close, determination of proteins engaged on the both sides of the
interaction can give us the insight to the mechanism of plant sub-
ordination to the parasite.
P1–13
Determination of chromosomal regions
affecting some production traits in F
2
intercross chickens
Z. Bulut
1
, E. Kurar
2
, Y. Ozsensoy
2
, M. Nizamlioglu
1
, M. Garip
3
,
A. Yilmaz
3
, T. Caglayan
3
, S. Dere
3
, V. Kurtoglu
4
and
M. Dogan
1
1
Faculty of Veterinary Medicine, Department of Biochemistry, Sel-
cuk University, Konya, TURKEY,
2
Faculty of Veterinary Medi-
cine, Department of Genetics, Selcuk University, Konya,
TURKEY,
3
Faculty of Veterinary Medicine, Department of Zoo-
technics, Selcuk University, Konya, TURKEY,
4
Faculty of Veteri-
nary Medicine, Department of Animal Nutrition, Selcuk
University, Konya, TURKEY
In this study, a F
2
level Denizli X White Leghorn population
(n = 441) was used to dissect chromosomal regions, which have
an affect on body weight and egg yield. DNA samples were iso-
lated from all F
0
,F
1
and F
2
animals using a standard phenol/
chloroform method. For chromosomal level screening, a total of
99 markers which were suitable for genome searching were ampli-
fied using Polymerase Chain Reaction (PCR). The resulting PCR
products were separated by capillary electrophoresis by using
Beckman Coulter CEQ-8000 Genetic Analysis System and geno-
types were identified at each marker loci. QTL Express Program
was used for QTL data analysis. In study, Quantitative Trait
Loci (QTL) were identified on chicken chromosome 1 (GGA1),
GGA2, GGA4, GGA8 and sex chromosome (GGAZ) for hatch-
ing weight, body weight at different ages and egg yield. There is
a need for narrowing these QTL regions by typing new markers
in these intervals and for identifying genes that have affect on
these economically important traits.
P1–14
Investigation of osmoprotection and cold
adaptation of halomonas halophilia DSMZ
4770 by proteomics
S. Ceylan
1
, B. Akbulut
1
, D. Kazan
1
and D. Kazan
2
1
Engineering Faculty, Bioengineering Department, Marmara
University, Istanbul, TURKEY,
2
TUBITAK Marmara Research
Center, Genetic Engineering and Biotechnology Institute, Kocaeli,
TURKEY
Halophiles are an important group of microorganisms that can
adapt to extremely saline environments. Since, they have the abil-
ity to function under extreme saline conditions, halophilic micro-
organisms are used at different industrial processes such as in the
production of protein/enzyme stabilizer metabolites (e.g. betain,
ectoin), in the removal of pollution in salinity effluents and in the
production of stable enzymes. Proteomics is the large-scale study
of proteins, particularly their structures and functions. Proteins
are vital parts of living organisms, as they are the main compo-
nents of the physiological metabolic pathways of cells. By under-
standing metabolic pathways of moderately halophilic
microorganisms it will be possible to modulate their metabolism
to enhance the production of their industrially important prod-
ucts. In this study osmoprotection and cold adaptation metabo-
lism of moderately halophilic Halomonas halophilia was
investigated. Halomonas halophilia was grown on different salt
concentrations (5% NaCL, -20% NaCl) and different tempera-
tures (19–37C). Growth at these conditions was investigated and
microorganisms were harvested at the end of exponential phase.
Afterwards, whole cell proteins of the DSMZ 4770 were
extracted and analyzed by proteomic tools. Two DE maps are
prepared according to tube gel systems with NEPHGE technique
in the first dimension and normal SDS-PAGE in the second
dimension. Protein profiles between normal and stress conditions
were determined and compared. Protein expression differences
were determined by MALDI-Tof Mass Spectrometer (Waters/Mi-
cromass). Proteins that involved in the osmoprotection and cold
adaptation were determined.
P1–15
Molecular typing of Staphylococcus aureus
strains from ovine mastitis by pulsed-field gel
electrophoresis and polymerase chain reaction
A. Ciftci
1
, E. E. Onuk
2
, A. Findik
1
, T. Yildirim
3
and
M. Unlu Sogut
4
1
Veterinary Faculty, Microbiology, Ondokuz Mayis University,
Samsun, TURKEY,
2
Veterinary Faculty, Diseases and Clinical
Sciences, Ondokuz Mayis University, Samsun, TURKEY,
3
Science Faculty, Biology, Amasya University, Amasya, TURKEY,
4
Medicine Faculty, Microbiology, Ondokuz Mayis University,
Samsun, TURKEY
Staphylococcus aureus is one of the most important etiological
agents of ovine mastitis. In order to develop effective control mea-
sures for mastitis, it is important to type S. aureus strains that
have considerable genetic heterogeneity. In the current study, 47
S. aureus strains isolated from ovine mastitis were typed by poly-
merase chain reaction (PCR) based on coagulase (coa) and protein
A (spa) polymorphisms and by pulsed-field gel electrophoresis
(PFGE). Eight different coa-types and four spa-types were identi-
fied by PCR. While the most prevalent coa-type was CG2
(42.56%), the spa-types, S4 and S1, were the most commonly
observed (44.68% and 38.29%, respectively). Nineteen different
pulsotypes were identified, and 12 of these were represented by a
single isolate. Pulsotypes J and K were predominant, and each
represented nine isolates (19.14%). All isolates belonging to J and
K pulsotypes were CG2. While all nine isolates belonging to J
pulsotype were S4, all isolates in K pulsotype were S1. Although
PFGE was found to be the best discriminatory technique for dis-
tinguishing strains, coa- and spa-types were found to be in correla-
tion with PFGE types and can be used for quick, preliminary
epidemiologic studies for detecting strains that may cause mastitis.
P1–16
Development and optimization of multiplex
polymerase chain reaction for identification
of Flavobacterium psychrophilum,Yersinia
ruckeri and Aeromonas salmonicida subsp.
salmonicida
E. E. Onuk
1
,A. Ciftci
2
, A. Findik
2
and Y. Durmaz
3
1
Veterinary Faculty, Fisheries and Fish Diseases, Ondokuz Mayis
University, Samsun, TURKEY,
2
Veterinary Faculty, Microbiology,
Ondokuz Mayis University, Samsun, TURKEY,
3
Veterinary Con-
trol and Research Enstitute, Fisheries and Fish Diseases, Samsun,
TURKEY
Bacterial cold water, enteric red mouth and frunculosis are the
common bacterial diseases of fish worldwide. The etiologic agents
Poster Presentations Abstracts
FEBS Journal 276 (Suppl. 1) 95–356 (2009) ª2009 The Authors Journal compilation ª2009 Federation of European Biochemical Societies 99