| Autor: |
Grava, Sandrine, Dumoulin, Pascal, Madania, Ammar, Tarassov, Ivan, Winsor, Barbara |
| Zdroj: |
Yeast; August 2000, Vol. 16 Issue: 11 p1025-1033, 9p |
| Abstrakt: |
We report here basic functional analysis of strains deleted for six open reading frames (ORFs), YNL059cand YNL148cfrom chromosome XIV and YOR145c, YOR152c, YOR161cand YOR162cfrom chromosome XV of Saccharomyces cerevisiae. ORFs were replaced with the KanMX4 resistance marker using a long flanking homology PCR strategy in FY1679 and W303 diploid strains. Replacement cassettes were constructed in plasmid pUG7 and the cognate wild‐type genes were recovered by gap repair. Sporulation and tetrad analysis revealed that deletion of YNL059c/ARP5was lethal for vegetative growth in strain W303 and caused severe growth defects in strain FY1679 while YOR145cwas essential for growth in both strains. Fusion of the green fluorescent protein (GFP) gene to the 3′ ends of the YNL059c/ARP5and YOR145ccoding sequences created functional chimeric genes at the respective chromosomal loci. Both Arp5–GFP and Yor145–GFP localized to the nucleus, Yor145–GFP concentrating in the nucleolus. The vectors containing the deletion cassettes and the cognate wild‐type genes, the oligonucleotides, and the deletant strains are available from the EUROFAN resource centre EUROSCARF (Frankfurt). Copyright © 2000 John Wiley & Sons, Ltd. |
| Databáze: |
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