Zbornik - Društvo genetičara Srbije

More documents

Recommendations

Info

50 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE II-Usm-3 Subotica, 30. novembar - 4. decembar 2004. ANALIZA PROMOTORSKOG REGIONA HUMANOG SOX3 GENA I IDENTIFIKACIJA TRANSKRIPCIONIH FAKTORA UKLJUÈENIH U REGULACIJU EKSPRESIJE Nataša Kovaèeviæ-Grujièiæ, Marija Mojsin, A. Krstiæ i Milena Stevanoviæ Institut za molekularnu genetiku i genetièko inenjerstvo, Beograd Familija SOX gena kodira proteine koji funkcionišu kao klasièni transkripcioni faktori i arhitektonske komponente hromatina i imaju znaèajnu ulogu u proliferaciji i diferencijaciji æelija tokom embriogeneze. Sox3 gen, smešten na X hromozomu, se eksprimira u centralnom nervnom sistemu u ranim fazama razviæa i predstavlja jedan od najranijih neuralnih markera kod kièmenjaka. Cilj našeg rada bio je definisanje i karakterizacija promotora SOX3 gena èoveka. Elongacijom prajmera reverznom transkripcijom odreðen je start transkripcije humanog SOX3 gena. Strukturno i funkcionalno je analiziran regulatorni region odgovoran za ekspresiju SOX3 gena u NT2/D1 æelijskoj liniji. Koristeæi promotor-reporterske konstrukte definisali smo minimalni promotorski region SOX3 gena odgovoran za njegovu bazalnu promotorsku aktivnost. PROMOTER ANALYSIS OF HUMAN SOX3 GENE AND IDENTIFICATION OF TRANSCRIPTION FACTORS IMPLICATED IN REGULATION OF ITS EXPRESSION Family of SOX genes encodes proteins that appear to govern cell fate decisions during embryogenesis by functioning both as classical transcription factors and architectural components of chromatin. Sox3, an X-linked member of the family, is expressed in the central nervous system from the earliest stages of development. It is considered to be one of the earliest neural markers in vertebrates playing the role in specifying neuronal fate. The aim of this study has been to determine and characterize the promoter of the human SOX3 gene and to elucidate molecular mechanisms underlying the regulation of its expression. Using primer extension method, we have identified the transcription start point and carried out the structural and functional analysis of the regulatory region responsible for SOX3 expression in NT2/D1 cell line. Using promoter-reporter constructs we have determined the minimal SOX3 promoter region that confers the basal promoter activity.
II-Usm-4 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 51 Subotica, 30. novembar - 4. decembar 2004. TIAZOFURIN NE DOVODI DO OŠTEÆENJA DNK U KOMET TESTU N. Ðeliæ 1 , Biljana Spremo-Potpareviæ 2 , Lada ivkoviæ 2 iV.Bajiæ 3 1 Katedra za biologiju, Fakultet veterinarske medicine, Univerzitet u Beogradu, Beograd 2 Institut za fiziologiju, Farmaceutski fakultet, Univerzitet u Beogradu, Beograd 3 Institut za biomedicinska istraivanja, «Galenka», Beograd Tiazofurin predstavlja nukleozid sa izraenim antitumorskim dejstvom. Intracelularno, tiazofurin se metabolièki konvertuje do tiazol-4-karboksamid adenin dinukleotida (TAD), analoga nikotinamid adenin dinukleotida (NAD), koji snano inhibira IMP dehidrogenazu. Usled inhibicije IMP dehidrogenaze kao kljuènog enzima u de novo sintezi purinskih nukleotida, dolazi do pada guanilata i na taj naèin se postie inhibicija rasta tumora kako in vitro,takoiin vivo. Cilj ovog rada bio je da se utvrdi da li tiazofurin moe da dovede do ošteæenja molekula DNK u humanim limfocitima, detektabilnih Komet testom (elektroforeza DNK pojedinaènih æelija, engl. single cell gel electrophoresis). Cela krv je izlagana eksperimentalnim koncentracijama (0.001, 0.002, 0.004 i 0.006 M) u trajanju od 30 min ili 2 sata na 37°C. Nakon tretmana i provere citotoksiènosti pomoæu tripan plavog, uzorci su centrifugirani, a zatim su æelije su resuspendovane u agarozi niske taèke topljenja, nanete na prethodno pripremljena predemetna stakla i ostavljena u rastvoru za liziranje na +4°C preko noæi. Sutradan je posle denaturacije DNK u puferu sa pH>13, izvršena elektroforeza u istom puferu na 0,7 V/cm. Nakon neutralizacije alkalija, predmetna stakla su odmah bojena etidijum bromidom i obavljena je analiza dobijenih kometa. Analizirano je taèno po 200 kometa za svaku eksperimentalnu koncentraciju. Statistièka analiza uraðena je neparametrijskim Wilcoxon-ovim testom. Tiazofurin u ispitanim koncentracijama nije ispoljio znaèajnu citotoksiènost (u svim uzorcima bilo je preko 90% vijabilnih limfocita). Pored toga, ne postoji statistièki znaèajno odstupanje u stepenu ošteæenja DNK u odnosu na kontrolne (netretirane) uzorke, kako pri tretmanu od 30 min, tako i pri tretmanu od 2 h. Na osnovu toga, moe se zakljuèiti da je rezultat testiranja tiazofurina u in vitro Komet testu negativan. TIAZOFURIN DO NOT INDUCE DNA DAMAGE IN THE COMET ASSAY Tiazofurin is a nucleoside with a profound antitumor effects. Tiazofurin is metabolically converted intracellularly to an analogue of nicotin amide adenin dinucelodide (NAD), tiazole-4-carboxamide adenine dinucleotide (TAD), a potent inhibitor of IMP dehydrogenase. Inhibition of this rate-limiting enyzme for de novo purine synthesis results in reduced levels of guanylates, inhibiting tumor growth in vitro and in vivo. The objective of the present investigation was to examine whether tiazofurin induces DNA damage in human lymphocytes detectable by single cell gel electrophoresis (Comet) assay. The whole peripheral venous blood was exposed to four experimental concentrations of tiazofurin (0.001, 0.002, 0.004 i 0.006 M) for either 30 min or 2 h at 37°C. After the treatment, we checked cytotoxicity using trypan blue dye exclusion. The samples were centrifugated, the cell pellet was resuspended in pre-heated low melting point agarose and layed on the precoated slides. Afterwards, the slides were left in lysing solution at +4°C overnight. Tomorrow, we performed DNA unwinding and electrophoresis in alkaline buffer (pH>13) at 0,7 V/cm. After the neutralisation of alkali, slides were stained with ethidium bromide and the comets were immediately scored. For each experimental concentration and exposure time we analysed exactly 200 comets. Statistical analyisis was performed by non-parametric Wilcoxon rank-sum test. Tiazofurin has not expressed significant cytotoxicity at any experimental point (lymphocyte viability was over 90%). Moreover, there is no significant departure in DNA damage
Page 1 and 2: DRUŠTVO GENETIČARA SRBIJE SERBIAN
Page 3 and 4: III KONGRES GENETIÈARA SRBIJE Subo
Page 5 and 6: Uvodno predavanje Opening lecture
Page 7 and 8: I tematska oblast / I topic: Geneti
Page 9: I-Uvo-2 ZBORNIK ABSTRAKATA III KONG
Page 12 and 13: 12 ZBORNIK ABSTRAKATA III KONGRESA
Page 43 and 44: II tematska oblast / II topic: Stru
Page 45 and 46: II-Uvo-2 ZBORNIK ABSTRAKATA III KON
Page 49: II-Usm-2 ZBORNIK ABSTRAKATA III KON
Page 53 and 54: II-Usm-6 ZBORNIK ABSTRAKATA III KON
Page 57 and 58: II-Pos-2 ZBORNIK ABSTRAKATA III KON
Page 63 and 64: III tematska oblast / III topic: Ge
Page 65: III-Uvo-2 ZBORNIK ABSTRAKATA III KO
Page 89 and 90: IV tematska oblast / IV topic: Ople
Page 91 and 92: IV-Usm-2 ZBORNIK ABSTRAKATA III KON
Page 99 and 100: IV-Usm-10 ZBORNIK ABSTRAKATA III KO
Page 101 and 102:
IV-Usm-12 ZBORNIK ABSTRAKATA III KO
Page 103 and 104:
Page 105 and 106:
Page 107 and 108:
Page 109 and 110:
Page 111 and 112:
Page 113 and 114:
Page 115 and 116:
Page 117 and 118:
IV tematska oblast / IV topic: Ople
Page 119 and 120:
IV-Pos-2 ZBORNIK ABSTRAKATA III KON
Page 121 and 122:
Page 123 and 124:
Page 125 and 126:
Page 127 and 128:
IV-Pos-10 ZBORNIK ABSTRAKATA III KO
Page 129 and 130:
Page 131 and 132:
Page 133 and 134:
Page 135 and 136:
Page 137 and 138:
Page 139 and 140:
Page 141 and 142:
Page 143 and 144:
Page 145 and 146:
Page 147 and 148:
Page 149 and 150:
Page 151 and 152:
Page 153 and 154:
Page 155 and 156:
Page 157 and 158:
Page 159 and 160:
Page 161 and 162:
Page 163 and 164:
Page 165 and 166:
Page 167 and 168:
Page 169 and 170:
Page 171 and 172:
Page 173 and 174:
Page 175 and 176:
Page 177 and 178:
Page 179 and 180:
V tematska oblast / V topic: Geneti
Page 181:
V-Uvo-2 ZBORNIK ABSTRAKATA III KONG
Page 184 and 185:
184 ZBORNIK ABSTRAKATA III KONGRESA
Page 186 and 187:
Page 188 and 189:
Page 190 and 191:
Page 192 and 193:
Page 194 and 195:
Page 196 and 197:
Page 198 and 199:
Page 200 and 201:
Page 203 and 204:
V tematska oblast / V topic: Geneti
Page 205 and 206:
V-Pos-2 ZBORNIK ABSTRAKATA III KONG
Page 207 and 208:
Page 209 and 210:
Page 211 and 212:
Page 213 and 214:
V-Pos-10 ZBORNIK ABSTRAKATA III KON
Page 215 and 216:
Page 217 and 218:
Page 219 and 220:
Page 221 and 222:
Page 223 and 224:
Page 225 and 226:
Page 227 and 228:
ZAVRŠNO PREDAVANJE CLOSING LECTURE
Page 229 and 230:
A Aksentijeviæ I. 194 Aleksiæ J.
Page 231 and 232:
Kovaèev L. 73, 90, 125, 140, 147 K
Page 233 and 234:
Stanojeviæ B. 223 Stanojeviæ J. 2
show all

Zbornik - Društvo genetičara Srbije

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?