Zbornik - Društvo genetičara Srbije

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78 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE III-Pos-7 Subotica, 30. novembar - 4. decembar 2004. KONSTRUKCIJA MOBILIZACIONOG VEKTORA ZA TRANSFER HETEROLOGIH GENA U LAKTOKOKAMA Maja Tolinaèki, Ivana Strahiniæ, Jelena Begoviæ, Lj. Topisiroviæ i M. Kojiæ Institut za molekularnu genetiku i genetièko inenjerstvo, Beograd Soj Lactococcus lactis subsp. lactis biovar. diacetylactis S50 je prirodni izolat poreklom iz maslaène maje. Soj S50 poseduje pet plazmida oznaèenih kao pS50-140, pS50-80, pS50-7a, pS50-7b i pS50-6. Odreðena je kompletna nukleotidna sekvenca dva mala plazmida pS50-7a i pS50-7b koji pokazuju 99% homologije meðu sobom (deo plazmida na kome se nalaze ORF1, ORF2, ORF3 i oriT pokazuje 100% identiènosti). Analizom nukleotidne sekvence pokazano je da plazmidi pS50-7a i pS50-7b pripadaju familiji laktokokalnih teta replicirajuæih plazmida. Zahvaljujuæi stabilnom odravanju unutar jedne æelije, kao i posedovanju mobilizacionih sposobnosti, plazmidi pS50-7a i pS50-7b predstavljaju dobru osnovu za konstrukciju mobilizacionog vektora uz pomoæ koga bi konjugacijom mogli biti preneti heterologi geni u netransformabilne sojeve. U cilju konstrukcije vektora za transfer heterologih gena iskorišæen je vektor za selekciju oridina replikacije pIS1. Za konstrukciju mobilizacionog vektora bilo je neophodno klonirati fragment poreklom iz plazmida pS50-7a, koji sadri esencijalne elemente za replikaciju, kao i oriT sekvencu, neophodnu in cis za transfer plazmida iz jedne æelije u drugu. Na osnovu analize nukleotidne sekvence utvrðeno je da EcoRI fragment, velièine 3392 bp, obuhvata minimalni replikacioni region kao i oriT sekvencu. Ovaj EcoRI fragment kloniran je u pIS1 vektor (konstrukt pISE3392) i uspešno eksprimiran u laktokokalnom soju VEL1122. CONSTRUCTION OF A MOBILIZABLE VECTOR FOR TRANSFER OF HETEROLOGUS GENES IN LACTOCOCCI Strain Lactococcus lactis subsp. lactis biovar. diacetylactis S50 is a natural isolate from starter for butter production. The strain S50 contains five different plasmids, designated as: pS50-140, pS50-80, pS50-7a, pS50-7b and pS50-6. The nucleotide sequence of both plasmids (pS50-7a and pS50-7b) was determined and data analysis revealed that the homology between them is 99%, (regions of plasmids containing ORF1, ORF2, ORF3 and oriT show 100% identity). Nucleotide sequence analysis of small plasmids (pS50-7a and pS50-7b) showed that they belong to a family of lactococcal theta replicons. The identification of mobilization ability of small plasmids and their stability in the cell would be benefical in the generation of mobilizable vectors for conjugative transfer of heterologus genes in non-transformable lactococci. For construction of mobilizable vector we used origin probe vector pIS1 (pUC19 derivate carring gene conferring resistence to erytromycin, and without replication ability in lactococci). The mobilizable vector shoud contain fragment from the plasmid pS50-7a, which carring essential elements for replication and oriT sequence (in cis acting sequence, necessary for plasmid transfer from one to another cell). The sequence analysis of EcoRI fragment of 3392 bp revealed the presence of oriT sequence and minimum region involved in plasmid replication. This EcoRI fragment was cloned into pIS1 vector (construct pISE3392) and successfully replicated in lactococcal strain VEL1122.
III-Pos-8 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 79 Subotica, 30. novembar - 4. decembar 2004. TESTIRANJE REGENERACIONE SPOSOBNOSTI INBRED LINIJA GAJENOG SUNCOKRETA Dragana Vasiæ, Ksenija Taški i D. Škoriæ Nauèni institut za ratarstvo i povrtarstvo, Novi Sad Sposobnost regeneracije in vitro je kod suncokreta (Helianthus annuus L.) ogranièena na samo mali broj genotipova. Kako je krajnji cilj kulture in vitro regeneracija biljaka, poeljno je testirati regneracionu sposobnost genotipova pre njihovog uvoðenja u eksperiment. Ispitivana je regeneraciona sposobnost 27 inbred linija gajenog suncokreta upotrebom metode Paterson i Everett (1985). Preæen je broj regenerisanih izdanaka iz eksplantata hipokotila. Genotipovi sa najveæim brojem regeneranata su dalje korišæeni u eksperimentima sa fuzijom protoplasta, kao roditelji recipijenti. TESTING OF REGENERATION CAPACITY OF CULTIVATED SUNFLOWER INBRED LINES In sunflower (Helianthus annuus L.), only small number of genotypes possesses a capacity to regenerate in vitro. As plant regeneration is the ultimate goal of in vitro culture it is necessary to test genotype regeneration capacity before its introduction into the experiment. Regeneration capacity of 27 cultivated sunflower inbred lines was tested using the method of Paterson and Everett (1985). Number of regenerated shoots from hypocotyl explants was observed. Genotypes with the highest number of regenerants were further used in the experiments with protoplast fusion as recipient parents.
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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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