Molecular Biology / Saccharomyces cerevisiae Yeast protocol protocols and methods

Technique / Molecular Biology / Saccharomyces cerevisiae Yeast protocol

Large-scale yeast transformation protocol
Protocols were developed by Petra Ross-Macdonald for Mike Snyder's LacZ fusion project at Yale University. (from YGAC Yale Genome Analysis Center) 1. Preparing the competent cells 2. Transforming yeast 3. Equipment required ...

Cloning by co-transformation and recombination in yeast new protocol
Cloning by co-transformation and recombination in yeast protocol. (Dept. Biochemistry, University of Washington) ...

Buffer medium recipes for fission yeast culture protocol
A buffer and medium recipe for fission yeast culture and other lab studies. ...

General yeast RNA protocols new protocol
Web supplementary materials (detailed method) for Genome-wide analysis of mRNA lengths in Saccharomyces cerevisiae. (Evan et.al, Standford University). Content: Total RNA preparation Isolation of poly(A) mRNA from total RNA using a Qiagen Oligote ...

Links to Yeast Protocols on the Web new recommended site
Yeast resources at Duke University. This web site provides comprehensive information about these genome collections, information and protocols for working with S. cerevisiae, links to important yeast related web sites and databases, and links to Duke labs that ...

Yeast Genetics, Biochemistry and Cell Biology site
An valuable site hosted by Herskowitz Lab. It includes lists of readings, links and original protocols on yeast genetics, cell biology and biochemistry. ...

Yeast nucleus staining troubleshooting
I am currently using DAPI to visualize yeast nuclei, but the signal from nucleus is often obscured by strong signal from mitochondria. So I am thinking about using Hoechst 33258 or 33342. Could someone give me some info what is the difference between ...

S. pombe work FAQ troubleshooting
This FAQ answers the most frequently asked questions about working with S. pombe. (from Forsburg lab at University of south california). ...

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09-May-2008 07:22 am

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Genome wide recruitment analysis of Rpb4, a subunit of Pol II in Saccharomyces cerevisiae reveals its involvement in transcription elongation.

Rpb4/Rpb7 subcomplex of yeast RNA polymerase II has counterparts in all multi-subunit RNA polymerases from archaebacteria to higher eukaryotes. The Rpb4/7 subcomplex in S. cerevisiae is unique in that it easily dissociates from the core, unlike in other organisms. The relative levels of Rpb4 and Rpb7 in yeasts affect differential gene expression and stress response. Rpb4 is nonessential in S. cerevisiae and affects expression of a small number of genes under normal growth conditions. Here, using ChIP on chip technique, we compared genome wide binding of Rpb4 to that of a core Pol II subunit, Rpb3. Our results showed that in spite of being nonessential for survival, Rpb4 was recruited on coding regions of most transcriptionally active genes similar to the core Pol II subunit, Rpb3, albeit to a lesser extent. The extent of Rpb4 recruitment increased with increasing gene length. We also observed Pol II lacking Rpb4 to be defective in transcribing long, GC rich transcription units suggesting a role for Rpb4 in transcription elongation. This role in transcription elongation was supported by the observed 6-azauracil sensitivity of rpb4Delta mutant. Unlike most phenotypes of rpb4Delta, the 6AU sensitivity of rpb4Delta strain was not rescued by over expression of RPB7. This report provides the first instance of a distinct role for Rpb4 in transcription, which is independent of its interacting partner, Rpb7.