Yeast electrophoresis

YEPD Yeast electrophoresis protein database (YEPD)... 

Joubert, R., Strub, J.-M., Zugmeyer, S., Kobi, D., Carte, N., Van Dorsselaer, A., Boucherie, H., and Jaquet-Gutfreund, L. 2001. Identification by mass spectrometry of two-dimensional gel electrophoresis-separated proteins extracted from lager brewing yeasts. Electrophoresis 22, 2969-2982. 

The principal molecular constituent of thin filaments is actin. Actin has been highly conserved during the course of evolution and is present in all eukaryotes, including single-celled organisms such as yeasts. Actin was first extracted and purified from skeletal muscle, where it forms the thin filaments of sarcomeres. It also is the main contractile protein of smooth muscle. Refined techniques for the detection of small amounts of actin (e.g., immunofluorescence microscopy, gel electrophoresis, and EM cytochemistry) subsequently confirmed the presence of actin in a great variety of nonmuscle cells. Muscle and nonmuscle actins are encoded by different genes and are isoforms. 

In E. Coli bacterial lysates, the proteome (i.e., the full array of proteins produced) was analyzed by isoelectric focusing and mass spectrometry.97 A comparison of capillary electrophoretic separation and slab gel separation of a recombinant monoclonal antibody demonstrated that the precision, robustness, speed, and ease-of-use of CE were superior.98 Seventy-five proteins from the yeast ribosome were analyzed and identified by capillary electrophoresis coupled with MS/MS tandem mass spectrometry.99 Heavy-chain C-terminal variants of the anti-tumor necrosis factor antibody DE7 have been separated on capillary isoelectric focusing.100 Isoforms differing by about 0.1 pi units represented antibodies with 0,1 or 2 C-terminal lysines. 

Three bacterial species (E. coli, P. putida, and S. rubidae) were separated on isoelectric focusing in methylcellulose coated capillaries, and three bacterial species (P. fluorescens, E. aerogenes, and M. luteus) and the yeast S. cerevisae, were separated by capillary electrophoresis in the presence of polyethylene oxide.101 The polymers served to minimize adsorption to the walls without causing cellular lysis. 

Breci, L., Hattrup, E., Keeler, M., Letarte, J., Johnson, R., Haynes, PA. (2005). Comprehensive proteomics in yeast using chromatographic fractionation, gas phase fractionation, protein gel electrophoresis, and isoelectric focusing. Proteomics 5, 2018-2028. 

L. Breci, E. Hattrup, M. Keeler, J. Letarte, R. Johnson, and P. A. Haynes. Comprehensive Proteomics in Yeast Using Chromatographic Fractionation, Gas Phase Fractionation, Protein Gel Electrophoresis, and Isoelectric Focusing. Proteomics, 5(2005) 2018-2028. 

Isolation of Plasmid Enriched DNA. The three strains of B. megaterium were cultured in yeast-soy broth (4% Bacto-yeast extract and 1.6% Bacto-soytone from Difco Laboratories, Detroit, pH 7.2). Plasmid DNA from each strain was isolated and purified by cesium chloride-ethidium bromide equilibrium density centrifugation following the procedure of Carlton and Brown ( ). Subsequent gel electrophoresis (0.5% agarose) revealed the presence of some chromosomal DNA contamination. 

Thus, the 1-naphthyl acetate activity from the cigarette beetle yeast does have properties, especially the solvent stability, that are likely to promote use for decontamination of lipophilic toxins. In addition, this activity (and in some cases additional molecular forms detected by gel electrophoresis) can be stimulated (based on higher activity relative to solvent controls) by malathion,... 

The biosynthesis in yeast of two enzymes that are D-mannoproteins has been studied. A membrane-associated isozyme of invertase (EC 3.2.1.26) has been shown to be a precursor of the external invertase.190 Its molecular weight, as determined by SDS-poly(acrylamide) gel electrophoresis, is 50,000, that is, smaller than that of the external invertase, and it correlates well with the presence of only the inner-core sugars of the final form. It is strictly bound to membranes, possibly those of the endoplasmic reticulum, and it can be completely split191 by endo-/3-N-acetylglucosaminidase H (EC 3.2.1.30). The addition of tunicamycin, which specifically inhibits formation of d-GIcNAc-PP-DoI, inhibits synthesis of external invertase, as well as further formation of the membrane-associated form, which completely disappears after addition of the antibiotic.190 In these aspects, the synthesis of this extracellular enzyme follows the pathway for secreted glycoproteins in animal systems. 

CM Ward, VC Trenerry, I Pant. The application of capillary electrophoresis to the determination of total niacin in concentrated yeast spreads. Food Chem 58 185-192, 1997. 

Schwartz, D. C., and C. R. Cantor, Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell 37 67-75, 1984. 

Contopolou, C.R., Cook, V.E., Mortimer, R.K. (1987). Analysis of DNA double-strand breakage and repair using orthogonal field alternation gel electrophoresis. Yeast 3,71-76. 

Fig. 4. Agarose gel electrophoresis of transcribed mRNA in vitro. Left lane DNA marker III digest. Middle and right lanes mRNA of yeast ubiquitin mRNA at...

Fig. 5. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) analysis of 15N-labeled yeast ubiquitin synthesized by wheat germ cell-free protein synthesis system. Left lane molecular weight markers. Middle lane yeast ubiquitin translation solution. Right lane wheat germ alone (control). Yeast ubiquitin is indicated by...

G21. Gross, D., Paper electrophoresis of the oligosaccharides synthesized from sucrose by yeast invertase. Nature 173, 487 (1954). 

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