Concept explainers
To analyze:
The result and describe the pathway that is followed by the protein by plotting a fluorescence intensity versus time graph with the help of the given table.
Given:
The green fluorescent protein (GFP) gene is fused with the gene of the viral protein. The normal cell is infected and the movement of the protein is monitored using fluorescence imaging equipment. The table shown below summarizes the result of the experiment:
Relative fluorescence intensity | ||||
Time (minutes) | Endoplasmic reticulum (ER) | Golgi | Cell membrane | Total |
0 | 0.95 | 0.05 | 0.00 | 1.00 |
20 | 0.64 | 0.28 | 0.08 | 1.00 |
40 | 0.38 | 0.39 | 0.23 | 1.00 |
60 | 0.17 | 0.38 | 0.44 | 0.99 |
80 | 0.05 | 0.28 | 0.65 | 0.98 |
100 | 0.00 | 0.25 | 0.70 | 0.95 |
150 | 0.00 | 0.05 | 0.77 | 0.82 |
200 | 0.00 | 0.00 | 0.75 | 0.75 |
Introduction:
The GFP is a protein that fluoresces and is obtained from the aquatic animal called jellyfish, having the scientific name Aequorea victoria. It is made up of 238 amino acids. It emits green fluorescent light in the presence of ultraviolet light. GFP is used widely in biological techniques. For example, GFP gene is fused with the gene of the viral protein. The normal cell is then infected and the movement of the protein is monitored using fluorescence imaging equipment. This method is used to track the pathway that is traced by the protein.
Explanation of Solution
The graph of fluorescence intensity against time for the protein that is present in the ER is given below:
This graph depicts the intensity of fluorescence against time for the protein that is present in the ER. Initially, the fluorescence intensity is high, which means that the protein is entering the ER directly after synthesis. The graph plot is gradually decreasing, which is showing that the fluorescence in the ER is decreasing and reaching zero. This means that the fluorescent protein has moved out of the ER.
The graph of fluorescence intensity against time for the protein that is present in the Golgi apparatus is given below:
This graph depicts the intensity of fluorescence against time for the protein that is present in the Golgi apparatus. Initially, the fluorescence intensity is increasing, which means that the protein is entering the Golgi after a considerable time. The graph plot is gradually decreasing, which is showing that the fluorescence in the Golgi is decreasing and reaching zero. This means that the fluorescent protein is also moving out of the Golgi.
The graph of fluorescence intensity against time for the protein that is present in the cell membrane is given below:
This graph depicts the intensity of fluorescence against time for the protein that is present in the cell membrane. The fluorescence intensity is increasing gradually and is maximized after some time, which means that the protein is embedded in the cell membrane.
The pathway that is followed by the protein is traced with the help of the graphs drawn. The protein enters the ER quickly after synthesis and then goes to the Golgi. From the Golgi, it moves to the cell membrane to form virus particles.
Therefore, the graphs are plotted and the pathway of the GFP fusion protein is traced with the help of the graphs. The pathway followed by the protein is from ER to Golgi and then to the cell membrane.
Want to see more full solutions like this?
Chapter 5 Solutions
Life: The Science of Biology
- protein is purified and at a concentration of 600 μg in 1.75 ml of buffer. You do an assay with your purified protein, and the assay requires 25 μg of biochemisfunase. What volume of the purified biochemisfunase would yield 25 μg? Show your calculation.arrow_forwardDraw the schematic diagram of the protein purification through hydrophobic column chromatography and explain the purpose of each step.arrow_forwardPlease describe the protein purification process with the aim of purifying a protein which locates on nucleus membrane.arrow_forward
- What are the example of nanostructures? Give at least ten and define each Note: [Do not copy on Google!]arrow_forward(MB = methylene blue) What is the concentration of the sample of the substance with the reporter absorbance of 0.40 at 595 nm?arrow_forwardDuring electrophoresis molecules are separated by three characteristics. What are they?arrow_forward
- Briefly compare the composition of DNA versus that of RNA. Answer the following questions pertaining to the experimental use of green fluorescent protein (GFP). What is the main advantage of GFP in terms of protein visualization in cells/organisms? Which microscope technique is used to visualize GFP-tagged proteins?arrow_forwardWhen you graph your protein assay data in Excel, should you include the absorbance of your unknown BSA sample? Explain why or why notarrow_forwardGive typing answer with explanation and conclusion Discuss in detail, the challenges of Protein A Purification. You may also compare how these challenges compare to Protein G Purificationarrow_forward
- Sketch the appearance after visualization of a protein mixture containing the seven proteins (fibrinogen, y-globulin, collagen, ovalbumin, myoglobin, hemoglobin, insulin) when subjected to two-dimensional (2D) gel electrophoresis.arrow_forwardGive only typing answer with explanation and conclusion You want to make 94 µL of the diluted Cell-Free extract for estimation of total protein. To do this, how much water will you add to the correct volume of the undiluted cell-free extract? Note: Cell-free extracts will need to be diluted 1:25 in water.arrow_forwardFirst, they ran samples of the enzyme on denaturing and non-denaturing gels; the results are shown in the figure to the right. In addition, they ran the protein through a calibrated gel filtration column, the results of which indicated that the PYC enzyme had a molecular weight of 540 kiloDaltons. In the figure, the rightmost column are where molecular weight standards of various sizes would occur if they’d been on the gels. How many subunits are in the PYC enzyme? Are the subunits the same, or are they different in some way? Are the subunits made up of distinct chains of primary structures, and, if so, how many? Are the chains the same, or are they different in some way?arrow_forward
- Human Anatomy & Physiology (11th Edition)BiologyISBN:9780134580999Author:Elaine N. Marieb, Katja N. HoehnPublisher:PEARSONBiology 2eBiologyISBN:9781947172517Author:Matthew Douglas, Jung Choi, Mary Ann ClarkPublisher:OpenStaxAnatomy & PhysiologyBiologyISBN:9781259398629Author:McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa StouterPublisher:Mcgraw Hill Education,
- Molecular Biology of the Cell (Sixth Edition)BiologyISBN:9780815344322Author:Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter WalterPublisher:W. W. Norton & CompanyLaboratory Manual For Human Anatomy & PhysiologyBiologyISBN:9781260159363Author:Martin, Terry R., Prentice-craver, CynthiaPublisher:McGraw-Hill Publishing Co.Inquiry Into Life (16th Edition)BiologyISBN:9781260231700Author:Sylvia S. Mader, Michael WindelspechtPublisher:McGraw Hill Education