Documents about Budding Yeast

  • 8 Pages

    Lecture2_08_Handouts

    Virginia Tech, BIOL 2104

    Excerpt: ... Lecture 2 August 28, 2008 Multicellular Organisms; Model Organisms in Cell and Molecular Biology; Tissue Culture Cells Multicellular organisms are composed by multiple cells. Differentiated cells contain the information necessary for the development of a complete organism 1 Model Organisms Molecular biologists have focused on E. coli The budding yeast , Saccharomyces cerevisiae, is the model organism for eucaryotes 2 Arabidopsis thaliana is the model organism for plant The fruit fly Drosophila melanogaster occupies a central place in biological research The nematode worm Caenorhabditis elegans is one of the most used model systems in developmental biology 3 The frog Xenopus leavis is a commonly used species in biochemistry labs X. tropicalis X. leavis Mammals are much more complex than other animals Why? The mouse is the model organism for mammals 4 However, most of what we know about human cell biology has come from studies of human cells Fibroblasts Myoblasts Oligodendrocyte prec ...

  • 4 Pages

    04

    UC Davis, MIC 170

    Excerpt: ... MIC170 4/9/09 Lecture 4 (Thursday, 4/9) Yeast Cell - 5. Cytoskeleton, continued A. Actin B. Tubulin Heterodimer of and tubulins (~50 kDa each) polymerize to form microtubules Fig. 12.42 from Textbook -> yeast tubulins have 75% identical amino acid sequence to human tubulins "dynamic instability" in living cells, each microtubule elongates and shrinks very rapidly due to continuous polymerization/depolymerization (movie) Fig. 12.44 from the textbook Microtubule Function: i) Forms spindle that separate chromosomes in mitosis (appears to be the most important function) ii) Nuclear positioning at the bud neck before mitosis - Fig. 2 in handout-KS3 1 MIC170 4/9/09 Yeast Life Cycle We'll look at the life cycle of the budding yeast S. cerevisiae (Fig. 4.19, handout-4) Developmental options in budding yeast A. Vegetative growth Haploid (DNA content = 1n) or diploid (2n) B. Stationary phase C. Mating - two haploid cells of different mating types (a and ) conjugate and form diploid D. Meiosis and sp ...

  • 4 Pages

    15

    UC Davis, MIC 170

    Excerpt: ... ing -correlates with initiation of DNA replication (G1S) DNA staining by fluorescent dye, such as DAPI (4',6'-diamino-2phenylindole) nucleus at the bud neck - G2 to early M nuclear division - M Budding yeast has no distinct G2 phase - mitotic spindle appears before DNA replication is completed(S phase) B. measure DNA content DNA flowcytometry using fluorescence-activated cell sorter (FACS) Fig. 4-31 (handout-8) stain DNA of cells with fluorescent dye that specifically binds to DNA and measure the amount of fluorescence for each cell (~10,000 cells/experiment) Fig. 17-6 (handout-8) Also informative to know the length of each cell cycle phases of growing culture - length of each cell cycle phase can be calculated from the data 2 MIC170 5/21/09 Two essential tools for cell cycle study in yeast A. Synchronized culture B. cdc mutants A. Synchronized culture (to analyze events specific to each cell cycle phase) If you just inoculate some yeast cells in liquid medium and incubate, y ...

  • 5 Pages

    16

    UC Davis, MIC 170

    Excerpt: ... MIC170 5/26/09 Lecture 16 (Tuesday, 5/26) Continued on "Cell cycle control" Two essential tools for cell cycle study in yeast A. Synchronized culture B. cdc mutants B. cdc mutants cell division cycle mutants-arrest at a specific point in the cell cycle Many temperature-sensitive (ts) cdc mutants were isolated both in budding yeast (Lee Hartwell) and fission yeast (Paul Nurse). (a) Isolation of cdc mutants i. isolate conditional (ts) mutants - select for mutants that can grow at the permissive temperature (25oC) but not at the restrictive temperature (37oC) mutants of interest are those that show a specific block in cell cycle; all cells in the culture arrest at one certain point in the cell cycle with certain cytological landmark e.g., all cells arrest without bud - cdc mutant that arrests in G1 ii. cdc mutant is deficient in a gene product required to get the cell past the specific point in the cell cycle Fig. 17-25 (handout-8) example: a cdc mutant defective in the initiation of S phase at the r ...

  • 6 Pages

    MCB 2007 November 14th Lecture Notes

    BC, BI 304

    Excerpt: ... s for cell-division cycle genes (cdc genes) o transformed yeast with every gene to identify which ones were responsible for cell cycle regulation o cdc2+ = wild type, cdc2- = wee phenotype (mutant, early entry into M phase) o code for the cdc2 protein o phenotype was easily observable in budding yeast s S. pombe and S. cerevisiae- cdc2 gene encodes for a CDK, cdc13 encodes a cyclin regulators: o cdc25 (decreases G2 length by stimulating transcription of cdc2) o wee1 (early entry into M via inhibition of cdc2 activity) regulatory genes are extremely conserved ...

  • 3 Pages

    midtermkey2

    UC Davis, MIC 170

    Excerpt: ... MIC170 Midterm May 5, 2009 Part I. Choose the ONE most appropriate answer for each of questions 1~5. 1. (2 points) The lipid bilayer of the plasma membrane is NOT permeable to glucose 2. (2 points) You are planning to construct a genomic library of the budding yeast S. cerevisiae. If the average size of the genomic DNA fragment in each plasmid is about 10 kb, you would want to make a library containing at least ( ) different plasmids to make sure the complete coverage of the entire genome. 6,000 3. (2 points) You are about to start chemical mutagenesis of the budding yeast S. cerevisiae. As you learned in the class, it would be ideal that a single mutation causes an interesting phenotype. Theoretically, to induce one mutation per cell in S. cerevisiae, the mutation frequency by mutagenesis needs to be about; 6x10-3/gene 4. (3 points) It's almost 9PM and you want to go home. You have an exponentially growing yeast cell culture, whose current concentration is 1.6 x 106 cells/ml. It has been determined that yea ...

  • 7 Pages

    03

    UC Davis, MIC 170

    Excerpt: ... MIC170 4/7/09 Lecture 3 (Tuesday, 4/7) Yeast cell 1. 2. 3. 4. 5. Cell surface Nucleus Mitochondria Vesicular organelles Cytoskeleton 2. Nucleus (continued from the last lecture) Essential elements in yeast linear chromosomes 1) CEN 2) ARS 3) TEL 1) CEN (centromere) sequences - One per chromosome (handout KS-2) = Sites of kinetochore formation - microtubules attach to kinetochore to separate sister chromatids in mitosis Fig. 5.19 from Textbook -> In budding yeast , 125-bp DNA is sufficient for centromere function <- many different proteins bind to form kinetochore Comparison of centromere DNA structures (Fig. 5.21 from Textbook) the 1 MIC170 4/7/09 2) ARS (Autonomously Replicating Sequence) - ~100bp First identified as sequences that enable plasmid DNA to replicate autonomously (Fig. 6.14 from Textbook) turned out it contains replication origin ORC (origin recognition complex), which contains six different proteins, binds to origin to initiate DNA replication Budding yea ...

  • 3 Pages

    test1key

    UC Davis, MIC 170

    Excerpt: ... tion of - and -( tubulin ) heterodimer. This characteristic of microtubules is known as "( dynamic instability )". G. In budding yeast , birth of daughter cells leaves distinct structures on the mother cell surface. They are called ( bud scar(s) ). 2. (5 points) Write "True" or "False" beside each of the following statements about yeast nucleus. True False Nucleolus is the site of ribosomal RNA synthesis and ribosomal assembly. Mitosis in yeast and fungi is known as "closed mitosis" because their nuclear envelope breaks down during mitosis. In comparison with the budding yeast Saccharomyces cerevisiae genome, the human genome contains ~267 times more genes. Each yeast chromosome has one ARS, the site of kinetochore formation. Every end of yeast chromosomes has a TEL sequence. These TEL sequences are synthesized by a specialized RNA polymerase called "telomerase". False False False 3. The mating type of the budding yeast S. cerevisiae is determined by the MAT locus on chromosome III. (a) (4 points) As we'l ...