Selective system based on fragments of the M1 virus for the yeast Saccharomyces cerevisiae transformation
Dmitri M. Muzaev , Andrey M. Rumyantsev , Ousama R. Al Shanaa , Elena V. Sambuk
Ecological Genetics ›› 2020, Vol. 18 ›› Issue (2) : 251 -263.
Selective system based on fragments of the M1 virus for the yeast Saccharomyces cerevisiae transformation
Background. A selective system based on the M1 virus of the yeast Saccharomyces cerevisiae was proposed.
Methods. To create a recipient strain, a DNA fragment encoding the killer toxin of the M1 virus under the control of the regulated promoter of the GAL1 gene was inserted into the genome of S. cerevisiae strains Y-1236 and Y-2177.
Results. Integration of such expression cassette leads to the conditional lethality – resulting strains die on a medium with galactose when killer toxin synthesis occurs. A linear DNA fragment containing the gene of interest flanked by sequences homologous to the promoter of the GAL1 gene and the termination region of the CYC1 gene is used to transform the obtained strains. During transformation due to homologous recombination, the sequence encoding the killer toxin is cleaved and the transformants grow on a medium with galactose.
Conclusion. The proposed selective system combines the main advantages of other systems: the use of simple media, without the need to add expensive antibiotics, and a simplified technique for constructing expression cassettes and selecting transformants.
yeast Saccharomyces cerevisiae / killer-toxins / M1 and M28 viruses / selective markers
Muzaev D.M., Rumyantsev A.M., Al Shanaa O.R., Sambuk E.V.
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