Zbornik - Društvo genetičara Srbije

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60 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE II-Pos-5 Subotica, 30. novembar - 4. decembar 2004. MUTACIONA ANALIZA POTENCIJALNOG MESTA VEZIVANJA RXRa PROTEINA U REGULATORNOM REGIONU SOX3 GENA Tijana Saviæ, Gordana Nikèeviæ i Milena Stevanoviæ Institut za molekularnu genetiku i genetièko inenjerstvo, Beograd SOX3 gen je èlan SOX familije gena koji ima znaèajnu ulogu u procesima neurogeneze i diferencijacije neurona. Poznato je da je ekspresija humanog SOX3 gena indukovana retinoiènom kiselinom (RA). U cilju definisanja elemenata odgovrnih za ovu indukciju napraviljena je serija delecionih konsrukata u okviru 5’ nekodirajuæeg regiona SOX3 gena. Funkcionalna analiza pokazala je da je najmanji od ovih konstrukata duine 31 bp zadrao sposobnost inducibilnosti RA. Na osnovu rezultata dobijenih u eksperimentima kotransfekcije sa ovim regionom zakljuèeno je da je dejstvo RA na ekspresiju SOX3 gena posredovano RXRa nuklearnim receptorom. U eksperimentima smanjene elektroforetske pokretljivosti pokazano je da se rekombinantni humani RXR protein vezuje za ovaj region. Mutacionom analizom ovog regiona tj. nizom eksperimenata smanjene elektroforetske pokretljivosti u kojima je poreðen afinitet vezivanja RXR nuklearnog receptora za mutirane probe u odnosu na wild type, definisan je RXR vezivni region. Analiza je pokazala da se ovaj vezivni region sastoji od 2 polumesta razdvojena sa 4bp. RXR vezivni region okarakterisan u ovom radu razlkuje se po svojoj sekvenci kao i po strukturi od RXR vezivnih mesta veæ opisanih u literaturi. MUTATION ANALYSIS OF THE POTENTIAL RXRa RESPONSE ELEMENT WITHIN REGULATORY REGION OF SOX3 GENE SOX3 gene is member of SOX gene family which has an important role in the process of neurogenesis and neuronal differentiation. It is known that the expression of the human SOX3 gene is induced by retinoic acid (RA). In order to deffine elements responsible for this induction a series of various deletion constracts of the 5’ noncoding region of the SOX3 gene has been made. Functional analysis has showed that the shortest of these constracts 31 bp in length is able to hold RA inducibility. Cotransfection experiments have indicated that the effect of RA on SOX3 gene expession is mediated by RXR nuclear receptor. Gelshift and supershift experiments have shown that the human recombinant RXR protein binds to this region. By comparation of the abiluty of RXR nuclear receptor to bind to wild type and serious of mutated oligonucleotides using gelshifts experiments, the RXR respnse region has been determined. Detailed analysis has shown that this response region consists of two halfsites separated with 4 bp. RXR response region, caracterised in this study differs by its sequence, as well as its structure, from already described RXR response elements.
II-Pos-6 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 61 Subotica, 30. novembar - 4. decembar 2004. NALIZA INTRONA U SOJEVIMA Bacillus spp. S. Stankoviæ 1 , V. Lazareviæ 2 , Blaenka Soldo 2 , Tanja Beriæ-Bjedov 1 , Jelena Kneeviæ-Vukèeviæ 1 i Draga Simiæ 1 1 Katedra za mikrobiologiju, Biološki fakultet, Univerzitet u Beogradu, Beograd 2 Départment de Microbiologie Fundamentale, Faculté de Biologie et de Médecine, Université de Laussane, Laussane, Suisse Kolekcija sojeva Bacillus spp. izolovanih iz prirodnih staništa je pretraivana pomoæu PCR na prisustvo umetnutih sekvenci (introna i intein-kodirajuæih sekvenci) u tandem genima bnrdE/bnrdF za ribonukleotid reduktazu SPâ profaga. Sekvenciranje PCR produkata je pokazalo tri nove konfiguracije u bnrdE-bnrdF tandemu koje se sastoje od introna grupe I u kodirajuæim regionima i ORF u intergenskom regionu. Sva tri soja pripadaju vrsti B. subtilis što je utvrðeno pomoæu API 50 CHB sistema za identifikaciju. Analiza genetièkih determinanti æelijskog zida, kao i inkorporacija radioaktivno obeleenog glicerola u æelijski zid, omoguæila je kategorizaciju osam do sada naðenih sojeva B. subtilis koji sadre umetnute sekvence u profagnim genima za ribunukleotid reduktazu u dve podvrste. Sojevi koji pripadaju subsp. subtilis sadre tri umetnute sekvence, introne grupe I i inteine, od kojih mnoge sadre ORF za pretpostavljenu «homing» endonukleazu. Sojevi koji pripadaju subsp. spizizenii sadre jedan ili dva introna grupe I koji ne sadre ORF. ANALYSIS OF Collection of Bacillus spp. strains isolated from natural habitats was screened by low stringency PCR for the presence of intervening sequences (introns and intein-coding sequences) in the SPâ prophage-related ribonucleotide reductase gene tandem bnrdE/bnrdF. The subsequent sequencing of the PCR products revealed three novel configurations within the bnrdE-bnrdF tandem based on the presence of the group I introns in protein coding sequences and an ORF in the bnrdE-bnrdF spacer. The three relevant strains belong to the species B. subtilis as determined by the API 50 CHB identification system. Analysis of the cell wall genetic determinants as well as the incorporation of radio-labeled glycerol into cell wall allowed categorization of eight B. subtilis strains known to contain intervening sequences in the prophage ribunucleotide reductase genes into two subspecies. Strains apparently belonging to the subsp. subtilis contain three intervening sequences, group I introns and inteins, many of which contain an ORF for the putative homing endonuclease. Strains of the subsp. spizizenii contain only one or two ORF-less group I introns.
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DRUŠTVO GENETIČARA SRBIJE SERBIAN
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III KONGRES GENETIÈARA SRBIJE Subo
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Uvodno predavanje Opening lecture
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I tematska oblast / I topic: Geneti
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ZAVRŠNO PREDAVANJE CLOSING LECTURE
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A Aksentijeviæ I. 194 Aleksiæ J.
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Kovaèev L. 73, 90, 125, 140, 147 K
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Stanojeviæ B. 223 Stanojeviæ J. 2
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Zbornik - Društvo genetičara Srbije

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