Novel methods are reported for evaluating and utilizing single chain fragment variable (scFv) antibodies derived from yeast-display libraries. Yeast-display was used to select scFv specific to invariant surface glycoproteins (ISG) of Trypanosoma brucei. A limiting step in the isolation of scFv from non-immune libraries is the conversion of highly active yeast-displayed scFv into soluble antibod...

Vesicular transport between successive compartments of the mammalian Golgi apparatus has recently been reconstituted in a cell-free system. In addition to ATP, transport requires both membrane-bound and cytosolic proteins. Here we report that the cytosol fraction from yeast will efficiently substitute for mammalian cytosol. Mammalian cytosol contains several distinct transport factors, which we...

Are plant cells just yeast cells with chloroplasts? Should plant cell biologists who don’t work on chloroplasts just switch to yeast to help solve the basic questions that are common to all eukaryotes? The recent completion of the sequencing of the entire yeast (Saccharomyces cerevisiae) genome and the availability of hundreds of well-characterized mutants certainly make this a tempting proposi...

The yield of fermentation process, bioethanol, is one the important product obtained from renewable energy sources. Yeasts are commonly used in ethanol production due to their high tolerance, productivity, and ability ferment different kinds simple sugars. Saccharomyces cerevisiae yeast species most industrial production. Therefore, isolation identification new strains with capacity very for bi...

Autolysis is a procedure frequently employed in the isolation of materials from yeast. Although the intracellular yeast proteinases play an important r81e in autolysis, these enzymes have not been characterized by using up to date methods of analysis, and the existing literature concerning yeast proteinases presents conflicting results. In 1917, Dernby (1) reported that yeast contains two prote...

The yeast three-hybrid system presents a valuable tool for detecting and analyzing RNA-protein interactions in vivo. A major drawback of the use of such a transcriptional reporter-based assay in a library screen is the frequent occurrence of false-positive results due to bait RNA-independent activation of the reporter gene. To minimize the isolation of false positives in three-hybrid library sc...

Ribosomal protein (rp) S6 is the major substrate of protein kinases in eukaryotic ribosomes. To facilitate the identification of cloned cDNAs for human rpS6, we used published amino acid (aa) sequence data for rat liver rpS6 and yeast (Saccharomyces carlsbergensis) rpS10 to design mixed oligodeoxynucleotide probes. Screening of several human cDNA libraries with these probes permitted the isolat...

Acetyl-CoA synthetase, utilized in a coupled reaction system, has been shown to be applicable to the spectrophotometric determination of propionic and methylmalonic acids in biological fluids. The isolation of acetyl-CoA synthetase from yeast is simpler than the purification from mammalian sources. This study also presents some properties of the yeast enzyme and compares it to the more extensiv...

Saccharomyces cerevisiae is an excellent model organism for the study of eukaryotic genetics. Easy manipulation of yeast DNA is essential to its role in research, and studies of gene expression or regulation require analysis of RNA. This chapter presents quick and straightforward methods to isolate genomic DNA, plasmid DNA, or RNA from yeast. The isolation protocols presented here, which utiliz...