Biology Lab report #1
The uptake of neutral red dye in a yeast cell using different solutions Abstract
Every cell transports materials in and out throught something called a membrane. There are many different methods of transport in the cell Saccharomyces cerevisiae (Serrano, 1977) We want to know does adding higher concentrations of azide more effectively block dye transport? We tested the transport of dye in yeast cells with a metabolic inhibitor. When we did this we showed no difference in the absorbance between different azide solutions, and our control. From this we concluded that azide has no effect on the transport through a yeast cell membrane. Introduction
Every cell has a layer of protection called the cell membrane. This cell membrane has many functions. The Saccharomyces cerevisiae cell has a selectively permeabile membrane which means it allows certian materials to pass through its membrane more or less than others ( Campbell et al., 2008). In a certian process called active transport these cells need energy to move material across the membrane ( Campbell et al., 2008). This energy that is being used is called adenosine triphosphate or ATP (Campbell et al., 2008). The production of ATP, which helps cells maintain a cells pH, helps with the uptake of neutral red dye because neutral red dye cannot be absorbed if the pH of the cell is reduced (Repetto, 2008). Saccharomyces cerevisiae has two types of active transporters through the cell membrane: primary and secondary active transporters (Stambuk, 2000). Sodium azide is a metobolic inhibitor which means it prevents ATP from being produced (Rowan University, 2009). In a study of Echerichia coli it was found that ATP was inhibited more than 90% using sodium azide ( Noumi, 1987).
The study of how things pass through different cell membranes is very common. So, studying how neutral red passes through a yeast cell is something that seems plausable to look at. We want to understand how...
Citations: 2009. Uptake of neutral red dye by Saccharomyces cerevisiae in the presence of a metabolic inhibitor. Biological Sciences, Rowan University. pp 1-8.
Campbell, N.A., Reece, J.B., Urry, L.A., Cain, M.L., Wasserman, S.A., Minorsky, P.V., Jackson, R.B. 2008. Biology, 8th ed. Pearson Benjamin Cummings, San Francisco CA, pp. 125-138.
Repetto, G., del Peso, A., LZurita, J.2008. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature Protocols. 3: 1125 - 1131.
Rikhvanov, E.G., Varakina, N.N., Rusaleva, T.M., Rachenko, E.I., Voinkov, V.K. 2002. The effects of sodium azide on the thermotolerance of the yeasts Saccharomyces cerevisiae and Candida albicans. Microbiology. 71(6): 662-665
Stambuk, B.U. 2000. A simple laboratory exercise illustrating active transportnext term in previous termyeast cellsnext term. Biochemistry and Molecular Biology Education. 28(6): 313-317.
Serrano, R. 1977. Energy requirments for maltose transport in yeast. European Journal of Biochemisrty. 80: 97-102
Noumi, T., Maeda, M., Futai, M. 1987. Mode of inhibition of sodium azide on H+-ATPase of Escherichia coli. Department of organic chemistry and biochemistry, the institute of scientific and industrial research, osaka university. 213(2): 381-384
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