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Yeast Transactivation Assay Lab Report

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This purpose of this experiment was to determine if a new yeast transactivation assay was able to determine estrogenic activity in estrogenic compounds. Throughout this experiment, researchers compared the detection ability of the new transactivation assay with the conventional transactivation assay. The new transactivation assay plasmid replaced the lacZ gene with GFP (green fluorescent protein) gene, and by doing so simplifies the process of determining estrogenic activity. The advantages of using GFP as the reporter instead of beta-galactosidase, which is the protein that the lacZ gene codes for, are numerous. The presence of beta-galactosidase is detected by photometric measurement, which requires the yeast cells to be disrupted which is both a time-consuming error and also a source of error. GFP is stable in a large pH range and is highly soluble, making it adaptable to a wide …show more content…

When 17beta-estradiol was added to the yeast cells, most emitted green fluorescence. There seemed to be a direct relationship with the magnitude of GFP expression and the concentration of 17beta-estradiol concentration. The results show that does GFP function as a satisfactory reporter gene for estrogen in this transactivation assay, satisfactory enough to replace the conventional beta-galactosidase transactivation assay. This study proves that GFP can replace beta-galactosidase as a reporter plasmid- a beneficial follow up study would be to see if there was a way to measure the level of enzyme activity in estrogenic compounds without transforming other genes upstream from the gene of interest, which in this case would be estrogen receptor alpha. If scientists can investigate ways to simply measure the activity of that expression plasmid, it would simplify this entire process and diminish the need for transactivation assays

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Macromolecule test 1 differs from the second chart by testing non-reducing sugars in the first test and proteins in the second. In depth the lab required to heat the sample at times, mix them, and add them to a warm water bath of 100 Celsius. The following graphs were obtained by following the guidelines within the

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In this experiment, one tested solutions that consisted of the standard yeast solution and added raw materials of interest. First, one measured and then added seven grams of Fleischmann 's Rapid-Rise yeast to a bottle containing two hundred and fifty milliliters of warm distilled water for the purposes of creating the standard yeast solution that was used in the fermentation experiment. Next, the standard yeast solution was added to each of the four fermentation flasks. After swirling the bottle, one and a half grams of the fermentation substrate was added into a beaker with fifteen milliliters of yeast suspension. The fermentation flask marked one required one and a half grams of Zulka Brand Morena Pure Cane Sugar as the fermentation substrate, the fermentation flask marked two required one and a half grams of Maseca Brand Corn Flour as the fermentation substrate, the fermentation flask marked three required one and a half grams of Carolina Biologicals Glucose as the fermentation substrate, and the fermentation flask marked four required one and a half grams of distilled water as the fermentation substrate.

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LABORATORY REPORT Activity: Enzyme Activity Name: Natalie Banc Instructor: Elizabeth Kraske Date: 09.26.2016 Predictions 1. Sucrase will have the greatest activity at pH 6 2. Sucrase will have the greatest activity at 50 °C (122 °F) 3.

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1153 Words | 5 Pages

This innovation has made it possible to relate physiological cell states to gene expression designs for considering tumors, progressions in disease, cell reaction to stimuli, proteins, metabolites, protein-protein collaborations, and modifications in a experiment. DNA microarrays can be utilized to identify DNA, or recognize RNA which is most regularly cDNA after reverse transcription that may be converted into proteins. This is done through Fluorescent colors. They are utilized to mark the extricated mRNAs or opened up cDNAs from the tissue or cell samples. The DNA array is then hybridized by incubating them overnight, and afterward washing to remove unimportant hybrids.

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984 Words | 4 Pages

With a new gene expresses, it can lead to genetic diversity. In daily basis experiments, scientists are introducing a new gene that can amplify the plasmid and produce large quantities of a new characteristic. This experiment builds a lot of beneficial for medicine and biotechnology. This technique called Transformation was introduced when a gene from the jellyfish was extracted and placed into the plasmid that encodes for Green Fluorescent Protein to produce the gene that give the characteristic of glowing green.

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529 Words | 3 Pages

Title: How Ph Levels Affected the Fermentation of Beer Hypothesis: The beer will be left with more sugar deposit as the Ph levels increase because alpha/beta -amylase will no longer function. Predictions: Alcohol Percentage Analysis for the Control and the Experimental During this experiment, the pH level was increased, therefore Alpha-Amylase was favored. Due to the nature of Alpha-Amylase cutting randomly through a large carbohydrate molecule, it leaves bigger sugars in the flask, which cannot be digested by yeast. Due to this, less reactions should occur in the experimental, therefore leading to a lower percentage of alcohol production, compared to the control.

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1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.

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The similarities that yeast share with humans are a main reason why yeast are a common experimental organism. In this experiment testing certain sugars on this organism can help understand how humans will react to these sugars. It is possible to make predictions as to how humans will react to these sugars by using yeast because of similar genes. Yeast are ideal experimental organisms because they are easy to get, and they are a simple organism (Yourgenome). In this experiment, four carbohydrates will be tested.

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778 Words | 4 Pages

What if a person were to learn that their perception of reality is wrong? Would that person defend their pride? Would they choose to accept the knowledge given? In Toni Cade Bambara's "The Lesson", a poverty-stricken girl is faced with the reality that she lives in a time where racial and economic inequalities exist. This short story tells how a young girl named Sylvia and her friends leave their neighborhood in the projects and take a field trip with an educated African American woman, Miss Moore, to a toy store in the rich part of the city.

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Plasmid DNA from E. coli cells was extracted and purified. To extract the plasmid DNA and purify it, the plasmid DNA had to be placed inside the bacterial cell through the process of transformation. The cells that took the plasmid DNA were grow grown in a media containing the antibiotics. Then the plasmid DNA is purified by removing genomic DNA, lipids and membrane proteins. With the DNA template containing the mutant sensor RNA, The DNA template was prepared for RNA synthesis using restriction endonuclease.

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To exclude technical variations, the target and reference genes were assayed on the same card. The cycle relative threshold method (Crt method) was applied. The expression of target genes across the samples was calculated using the equation ΔCrt, in which [ΔCrt = target gene Crt – the mean of reference genes Crt]. A lower ΔCrt indicates higher gene

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780 Words | 4 Pages

The Effect of Sugar Concentration on CO2 Production by Cellular Respiration in Yeast Introduction In this lab, our main focus was to find how sugar concentration affect yeast respiration rates. This was to simulate the process of cellular respiration. Cellular respiration is the process that cells use to transfer energy from the organic molecules in food to ATP (Adenosine Tri-Phosphate). Glucose, CO2, and yeast (used as a catalyst in this experiment) are a few of the many vital components that contribute to cellular respiration.

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451 Words | 2 Pages

Yeast is alive because it can to metabolize and respond to environmental changes. The purpose of the first experiment was to determine whether yeast can metabolize. The bromothymol blue solution with yeast changed from blue to yellow. Bromothymol blue is an acid-base indicator that turns yellow in the presence of acid. The color change indicates that carbonic acid was formed from the reaction of water and carbon dioxide, a byproduct of metabolization.

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1000 Words | 4 Pages

Cell viability assay: Introduction. Methods in Molecular Biology 740: 1-6. ThermoFisher Scientific. [Internet].

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1371 Words | 6 Pages

Biochemical tests are the tests used for the identification of bacterial species based on the differences in the biochemical activities of different bacteria. Bacterial physiology differs from one species to the other. These differences in carbohydrate metabolism, protein metabolism, fat metabolism, production of certain enzymes and ability to utilize a particular compound help them to be identified by the biochemical tests. Gram’s stain was originally devised by histologist Hans Christian Gram in 1884. Gram-positive bacteria stain purple, while Gram-negative bacteria stain pink when subjected to Gram staining.

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