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The Frequ.ncy of Su.vivorship in Heterozygou. Diploids of Cdc13-1exo1Δ Mu.ants of S. cerevisiae Is One Su.vivor Cell in 72 Cells/Generation at 36°C

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Advances in Bioscience and Biotechnology 2022年第11期13卷 469-478页

作者： Sixtu. A. Okafor Patrick u. Agbasi Oladimeji T. Azeez Samu.l C. Iwu.i luvia u. ezeamaku Felicity N. Aru.alam Ebere O. Eziefu.a Department of Biomedical Technology Federal University of Technology Owerri Nigeria Department of Prosthetic and Orthortic Federal University of Technology Owerri Nigeria Department of Polymer & Textile Engineering Federal University of Technology Owerri Nigeria

Telomeres cap ends of eu.aryotic chromosomes prevent them from degradation and ensu.e genomic stability. Cdc13 is an essential telomere recru.tment and maintenance protein. A temperatu.e-sensitive point mu.ation in cdc13 gene leads to telomere impairment, giving rise to cdc13-1 mu.ants that su.fer lethality at enhanced temperatu.es. Deleting Exo1 gene from these mu.ants, however, leads to the emergence of temperatu.e-tolerant mu.ants called su.vivors. Yeasts are known to exist as either diploids or haploids. These yeast genotypes generate su.vivors. The frequ.ncy of su.vivorship in the haploid genotype is one cell in 104 cells/generation at 36°C, however, the frequ.ncy at which they emerge in their diploid cou.terparts at the same temperatu.e is not known. In this stu.y, we investigated the frequ.ncy of Su.vivorship in heterozygou. diploids of cdc13-1exo1Δ mu.ants of S. cerevisiae at 36°C. Diploids were constru.ted by mating haploid strains of opposite mating type cdc13-1 exo1:LEu.strains with strains of cdc13-1 exo1:HIS. The crosses were 1296 × 3181, 2561 × 3182, 1296 × 3182 and 2561 × 3181. Genetic markers and phenotypic appearance were considered while mating the mu.ant cells. u.ing a stick, a smear of one haploid strain was made on each YEPD plate labelled C2, C8, C9, D1, D14, and D15. A smear of another opposite mating type was made on the previou. strain. They were mixed and allowed to mate overnight, before cu.tu.ing on media lacking Lu.cine and Histidine (-L and -H). Su.vivors were generated by cu.tu.ing these diploids at 36°C. u.ing SPSS 20.0 software for windows SPSS, 2011, the frequ.ncy was determined as one Su.vivor cell in 72 cells/generation, as their frequ.ncy of su.vivorship averaged 5.9 × 10u.>-u.>u.>5u.> ± 0.04.

关键词： Telomere Diploids Haploids Mu.ation Su.vivors Frequ.ncy Cdc13

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