Disruption of six Saccharomyces cerevisiae ORFs on chromosome XII results in three lethal disruptants.
نویسندگان
چکیده
Six ORFs of unknown function from the left arm of chromosome XII of Saccharomyces cerevisiae were chosen for a reverse genetic approach to provide materials to assist in assignment of function. A two-step PCR using long-flanking homology was employed to amplify disruption cassettes consisting of a kanMX gene as selectable marker flanked by 250-350 bp long regions homologous to the target gene. The diploid strains FY1679 and CEN.PK2 were transformed with the replacement cassettes and transformants were selected for geneticin (G418) resistance. Correct targeting of the replacement cassettes at the genomic locus was verified by Southern blot analysis with the kanMX gene as a probe. Disruption cassettes were cloned in pUG7 plasmid for systematic gene inactivation in other yeast strains and the cognate genes were cloned in pRS416 plasmid for gene complementation studies. Sporulation and tetrad analysis of heterozygous disruptants showed that three of the six ORFs [YLR141w (RRN5), YLR145w and YLR147c (SMD3)] were essential genes that were complemented by their cognate genes. ylr146c Delta (spe4) homozygous diploids showed enhanced sporulation efficiency, whereas ylr147c Delta heterozygous diploids failed to sporulate in the FY1679 but not in the CEN.PK2 genetic background. The other two disruptants [ylr143w and ylr144c (acf2)] gave no phenotype.
منابع مشابه
Initiation of Ageing Process by Meiotic and Mitotic Recombination within the Ribosomal DNA Genes in Saccharomyces cerevisiae
In the budding yeast of Saccharomyces cerevisiae the tandem repeated of rDNA genes are located onchromosome XII, which is in the nucleolus. There are different types of proteins in the nucleoluskeleton,silencing proteins have got important role in nucleolus.It is shown that meiotic recombination between nonsister chromatids in the rDNA genes are stronglysuppressed, and s...
متن کاملThe nucleotide sequence of Saccharomyces cerevisiae chromosome IV.
The complete DNA sequence of the yeast Saccharomyces cerevisiae chromosome IV has been determined. Apart from chromosome XII, which contains the 1-2 Mb rDNA cluster, chromosome IV is the longest S. cerevisiae chromosome. It was split into three parts, which were sequenced by a consortium from the European Community, the Sanger Centre, and groups from St Louis and Stanford in the United States. ...
متن کاملGenome Organization of the Three Identical ATP1 Genes on the Left Arm of Chromosome II of Saccharomyces cerevisiae: Sequence Analysis of the 35-kb Region Containing Three ATP1 Genes
The ATP1 gene (YBL099w) encoding the F1F0-ATPase complex subunit of Saccharomyces cerevisiae is present on the left arm close to the telomere of chromosome II, and only one copy was reported by the Genome Project. Recently, we reported that three ATP1s designated ATP1a, ATP1b and ATP1c are located on chromosome II, with different distances between ATP1a and ATP1b, and ATP1b and ATP1c as identif...
متن کاملEffects of bioencapsulated Daphnia magna with Saccharomyces cerevisiae on the growth and feeding performance of Persian sturgeon (Acipenser persicus) larvae
BACKGROUNDS: Optimization of microbial compositions and load in live food during the process of bioencapsulation is one of the most important concerns in aquaculture, as it can promote the growth and feeding parameters of fish larvae. OBJECTIVES: The aim of this study was to determine the growth and feeding performance of Persian sturgeon larvae fed with bioencapsulated D. magna with Saccharomy...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Yeast
دوره 19 1 شماره
صفحات -
تاریخ انتشار 2002