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Registration Number

0225U004216, (0124U004341) , R & D reports

Title

Study of the mechanisms of action of new genes involved in the regulation of riboflavin synthesis in the flavinogenic yeast Candida famata

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Клонування, делеція та надекспресія диких та виявлених у мутанта AF-4 мутованих алелей генів SEF2, MET2 та RIB3 у штамі C. famata дикого типу та у мутантів sef1Δ та sfu1Δ.

Head

Fedorovych Dariya V., д.б.н.

Registration Date

26-11-2025

Organization

Institute of Cell Biology of the National Academy of Sciences of Ukraine

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To investigate the interaction of the previously identified transcription factor Sef1 with the promoters of the structural genes of riboflavin synthesis. To identify new genes involved in the regulation of riboflavin biosynthesis in the flavinogenic yeast Candida famata and to study their mechanisms of action by cloning and modifying them. To study the phenotypes of the obtained strains, namely, the accumulation of riboflavin, the activity of enzymes and the expression of the corresponding structural genes, as well as the content of iron in the cells.

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Object of research: regulation of riboflavin biosynthesis in the flavinogenic yeast Candida famata Research goal: study the mechanism of action of new genes involved in the regulation of riboflavin biosynthesis in the yeast C.f amata. Goal of the stage: to identify new genes involved in the regulation of riboflavin biosynthesis in the flavinogenic yeast C. famata and to study the mechanisms of their action by cloning and modification. Research methods: bioinformatic, genetic engineering, microbiological, biochemical. Bioinformatic work was carried out to search for and analyze the transcription factor Sef2, the MET2 gene, and the structural gene for riboflavin biosynthesis RIB3 in the yeast C. famata. Integrative vectors containing native or mutated alleles of the SEF2, MET2 and RIB3 genes of C, famata found in the AF-4 mutant were constructed under the control of their own promoters or a strong constitutive promoter of the TEF1 gene, which encodes a translation elongation factor. The constructed plasmids were used to obtain transformants with overexpressed SEF2, MET2 and RIB3 genes based on the wild-type strain C, famata VKM Y-9, also a riboflavin producer of AF-4. Cassettes for deletion of the SEF2 and MET2 genes were constructed and used to obtain strains with deleted sef2Δ and met2Δ genes C, famata. It has been established that the transcription factor Sef2 and the product of the MET2 gene are involved in the regulation of riboflavin biosynthesis in the yeast C. famata, Key words: Riboflavin biosynthesis, genetic engineering, yeast Candida famata, transcription factor Sef2, RIB3 and MET2 genes,

Product Description

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Lubov R. Fayura
Lubov S. Dzanaeva
Andriy O. Tsyrulnyk
Romanov Serhii O.

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2025-11-26