Your bacterial paints ("Bio-paints") are transformed E. coli that contain genes for producing β-lactamase, an enzyme that degrades the antibiotic ampicillin. The presence of this enzyme allows your Bio- paints to do which of the following? Produce GFP (Green Fluorescent Protein) Grow in the presence of ampicillin Grow in the presence of all antibiotics Produce GFP (Glowing Flavor Paint)
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Step 1: The bacteria have been transformed to produce β-lactamase, an enzyme that degrades ampicillin. Show more…
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To determine whether bacterial cells are competent, cells are grown with external DNA carrying two genes: green fluorescent protein and ampicillin resistance. Then, they are plated on nutrient agar with and without ampicillin. Plates are examined for fluorescent colonies under UV light after incubation. Indicate the expected results if cells are competent. a. There will be no growth on either type of plate b. Fluorescent colonies will be present on the ampicillin agar plates, but there will be no growth on the nutrient agar plates c. Fluorescent colonies will be present on both types of plates d. Colonies will be present on both types of plates, but will only fluoresce on the ampicillin agar plates
Adi S.
Plasmid Diagram The plasmid that we will introduce into E. coli bacterial cells contains two genes – the gene for the green fluorescent protein and an ampicillin resistance gene (see diagram above). Ampicillin is an antibiotic, and the amp resistance gene allows us to select for bacterial cells that have taken up the plasmid and express the ampicillin resistance gene. The green fluorescent protein (GFP), is a protein originally identified in jellyfish, which is inherently fluorescent. Cells expressing GFP are autonomously fluorescent and can be visualized through the use of a fluorescent microscope. Furthermore, if the gene for GFP is incorporated into cells of other species, these cells also become autonomously fluorescent. This provides researchers with a means to visualize cells shape and structure in live cells, as well as monitor shape changes of these cells over time. In addition, using molecular biology techniques, the gene for GFP can be fused to the gene of another protein. This allows researchers to then investigate the subcellular localization of this protein in live cells (see localization of GFP-Tubulin in the figure above). Given these properties, GFP (and other color derivatives) have become a valuable tools for cell biology, gene expression, developmental biology, and numerous other areas of biological research. This lab involves the incorporation of the gene for GFP into bacterial cells and then visualizing the expression of GFP in cells that have been successfully transformed with this gene. (Q3) What is the benefit of using GFP located within a plasmid for researchers?
Suppose you transformed Escherichia coli with the pGLO plasmid, which contains the gene for the GFP protein and the gene that makes E. coli ampicillin-resistant. In your transformation plate, upon exposure to UV light, you saw mixtures of green and white colonies. You suspect that the antibiotic is not working properly. Mark ALL that apply about the components of the petri dish you will use to assess the antibiotic activity: E. coli + pGLO, E. coli - pGLO, LB + ampicillin, LB - ampicillin, arabinose must be present, arabinose must be absent, arabinose will not change the outcome for this petri dish.
Madhur L.
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