SVIBOR - Project code: 1-08-076

MINISTRY OF SCIENCE AND TECHNOLOGY

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SVIBOR

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Project code: 1-08-076


Recombination in yeast - substrate and genetic control


Main researcher: ALAČEVIĆ, MARIJA (182)


Assistants
Type of research: basic
Duration from: 03/01/91. to 03/01/94.
Papers on project (total): 32
Papers on project quoted in Current Contents: 9
Institution name: Prehrambeno-biotehnološki fakultet, Zagreb (58)
Department/Institute: Department of Biochemical Engineering Laboratory of Biology and Microbial Genetics
Address: Pierottijeva 6
City: 10000 - Zagreb, Croatia
Communication
Phone: 385 (041) 417-044
Fax: 385 (041) 411-436
E-mail: zgazo@mapbf.pbfrng.hr
Summary: Hybrid DNA is the central intermediate in homologous recombination. Genetic studies have revealed that it may also contain unpaired or mismatched bases, in which case it is called heteroduplex DNA. We are studying the influence of such heterologies on the efficiency of homologous recombination using plasmid (both double and single-stranded) integration into the yeast chromosome as a model system. Plasmids with changed bases are constructed by in vitro mutagenesis, while the ratio betweeen homologous and homoeologous integrations is determined by Southern blot technique. The effect of UV-induced lesions on transformation with single- and double-stranded integrative plasmids was also investigated, as well as the feasibility of yeast transformation with single-stranded linear DNA. We also studied the influence of mismatched bases on homologous recombination in Escherichia coli.

Keywords: homologous recombination, DNA heteroduplex, integrative plasmids, illegitimate integration, single-stranded DNA, Saccharomyces cerevisiae

Research goals: Recombinational events are usually described as either homologous or illegitimate. In the former case, the recombining molecules share the same, or almost same sequence of nucleotides, while in the lather case the recombining molecules are heterologous or they share only very short homology. In our research we investigate the role of heterologies on homologous recombination in yeast Saccharomyces cerevisiae, using transformation with non-replicative plasmid as a model system. This research will be complemented by the research performed at the level of yeast and bacterial genome. We want to find out how much divergency is tolerated in the process of homologous recombination and to describe the mechanisms that prevent recombination between diverged sequences. We will also investigate transformation with UV-irradiated plasmids and the possibility of illegitimate plasmid integration in yeast. In addition to its biological interest this investigation could also lead to the development of alternative strategies for yeast strain construction.

COOPERATION - PROJECTS


  1. Name of project: CI1-0528-M DNA repair and strain construction in yeast
    Name of institution: Institut Curie, Institut J. Monod
    City: Pariz, Francuska

COOPERATION - INSTITUTIONS


  1. Name of institution: Institut Jaques Monod
    Type of institution: University/Faculty
    Type of cooperation: Joint project
    City: Pariz, Francuska

  2. Name of institution: Institut Curie
    Type of institution: State institute
    Type of cooperation: Systematic exchange of information
    City: Pariz, Francuska
Other information about the project.
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Last update: 10/16/95
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