Pearson eText Biology: Life on Earth with Physiology -- Instant Access (Pearson+)
12th Edition
ISBN: 9780135755785
Author: Gerald Audesirk, Teresa Audesirk
Publisher: PEARSON+
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Chapter 3.4, Problem 1CSC
Infectious prions such as those that cause mad cow disease are abnormally folded versions of a protein that is found throughout the body. The secondary structure of the normal prion protein is primarily helical. Infectious prions, however, fold into pleated sheets. The pleated sheets are so stable they are unaffected by the enzymes that break down normal prion protein. As a result, infectious prions accumulate destructively in the brain.
Helices and pleated sheets are the two major secondary structures of proteins. What do protein tertiary and Quaternary structures look like?
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1)Which plate did you see purple/pink/blue bacterial cells? Why did you see this growth? Explain your answer in terms of transformation and plasmids?
2) Calculating Transformation Efficiency
For the +DNA/+Amp/+IPTG plate, record the following:
Number of transformants (colonies): _________________
Nanograms of plasmid DNA added: 50 ng
Final recovery volume: 0.50 mL
Volume plated: 0.25 mL
Transformation efficiency equation:
Transformation efficiency = Number of transformants / µg of DNA x Final volume at recovery (mL)/ volume plated (mL)
3) Using the equation above, calculate the transformation efficiency.
4) Describe the success of the transformation efficiency of this demo based on the calculation you did above?
1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results?
2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab?
3) Why are the cells incubated at 42°C?
Overview of Transformation Protocol
-Prepare competent bacteria for transformation:
Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.
Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.
-Transformation procedure:
Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.
Add CaCl2 to both tubes.
Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.
Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.
Add recovery broth to both tubes.
Incubate both tubes in a 37 C water bath for 5 minutes.
Questions:
1) What differences would you expect to see between the…
Chapter 3 Solutions
Pearson eText Biology: Life on Earth with Physiology -- Instant Access (Pearson+)
Ch. 3.1 - which of these is/are polar molecules? (you may...Ch. 3.1 - define organic molecules and explain why carbon is...Ch. 3.1 - explain why functional groups are important in...Ch. 3.1 - name and describe the properties of seven...Ch. 3.2 - define organic molecules and explain why carbon is...Ch. 3.3 - Describe hydrolysis of this molecule.Ch. 3.3 - describe the major types of carbohydrates?Ch. 3.3 - provide examples of each type of carbohydrate and...Ch. 3.4 - Look up the rest of the amino acids and. based on...Ch. 3.4 - Infectious prions such as those that cause mad cow...
Ch. 3.4 - Why do many proteins, when heated excessively....Ch. 3.4 - Why a Perm Is (Temporarily) Permanent?Ch. 3.4 - describe protein subunits and how proteins are...Ch. 3.4 - explain the four levels of protein structure and...Ch. 3.4 - list several functions of proteins and provide...Ch. 3.4 - Prob. 4CYLCh. 3.5 - describe the general structure of nucleotides?Ch. 3.5 - list three different functions of nucleotides?Ch. 3.5 - explain how nucleic acids are synthesized?Ch. 3.5 - give two examples of nucleic acids and their...Ch. 3.5 - Puzzling Proteins All cells use DNA as a blueprint...Ch. 3.6 - What kind of reaction breaks this molecule apart?Ch. 3.6 - An obese 55-year-old woman consults her physician...Ch. 3.6 - Why are steroid hormones able to diffuse through...Ch. 3.6 - compare and contrast the structure and synthesis...Ch. 3.6 - describe the functions of fats, oils, and waxes?Ch. 3.6 - Prob. 3CYLCh. 3.6 - Prob. 1CTCh. 3 - Polar molecules a. dissolve in lipids. b. are...Ch. 3 - Prob. 2MCCh. 3 - Prob. 3MCCh. 3 - Which of the following is not composed of...Ch. 3 - Prob. 5MCCh. 3 - In organic molecules made of chains of subunits,...Ch. 3 - Prob. 2FIBCh. 3 - Prob. 3FIBCh. 3 - Prob. 4FIBCh. 3 - Fill in the following with the appropriate type of...Ch. 3 - Prob. 1RQCh. 3 - List the four principal classes of biological...Ch. 3 - Prob. 3RQCh. 3 - Prob. 4RQCh. 3 - Prob. 5RQCh. 3 - Describe the synthesis of a protein from amino...Ch. 3 - Where in nature do we find cellulose? Where do we...Ch. 3 - Based on their structure, sketch and explain how...Ch. 3 - Prob. 2ACCh. 3 - Prob. 3AC
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- Overview of Transformation Protocol -Prepare competent bacteria for transformation: Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure. Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed. -Transformation procedure: Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA. Add CaCl2 to both tubes. Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube. Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes. Add recovery broth to both tubes. Incubate both tubes in a 37 C water bath for 5 minutes. Questions: 1)What is the selectable marker in this experiment? How…arrow_forwardBased on your results, which suspect's DNA best matches the DNA found at the crime scene?arrow_forwardIn oxidase test with Pseudomonas aeruginosa, the cell cultures on the slide turn colorless to be purple after tetra-methyl-p-phenylenediamine dihydrochloride (TMPD) is added. In the reaction, OTMPD is electron acceptor O cytochrome c is the electron source oxygen is terminal electron acceptor OH2 produced is electron donorarrow_forward
- You will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. You have shipped your samples off for sequencing and are now waiting for the results. Out of curiosity (and maybe boredom...) you decide to test your culture for the Catalase and Oxidase enzymes. Upon testing your sample for catalase, you don't see any bubbles; however, you do see a color change to purple during the Oxidase test. What results can you conclude from this? O Catalase-/ Oxidase + O Catalase +/ Oxidase + Catalase + / Oxidase- O Catalase / Oxidase - O None of the abovearrow_forwardWhich of the following is not a strength of using 16S rRNA for phylogenetic analyses? OA. It's cheap OB. It's easy to do C. It can be used to identify all the way down to the strain level OD. Both A & B OE. None of the abovearrow_forwardWhy are molecular approaches important to the field of microbial taxonomy and phylogeny? Phylogenetic inferences based on molecular approaches provide the most robust analysis of microbial evolution currently available. It allows for the collection of a large and accurate dataset from many organisms Almost no fossil record was left by microbes when compared to plants and animals All of the above None of the abovearrow_forward
- You will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. You have already cultured it and gone through the plate isolation procedure. Before you ship your samples off for sequencing, you want to do one final check of the A260 ratios. You get back the following ratios: A260/280 ratio is 1.89; A260/230 is 2.01. These ratios are close enough to the accepted "pure" values so they could be considered "pure" and mostly (if not completely) free of contaminants from the PCR process. True Falsearrow_forwardYou will use the following scenario to answer a group of 5 questions. You have isolated a microbe from an environmental sample. The microbe has the ability to perform a new metabolic reaction at a very low temperature, so you are excited that it could be a new species. After receiving your sequence back from the sequencing lab, you feel that you have, in fact, discovered and isolated a new species. You ask a fellow labmate about how you should proceed, and he tells you the following is the proper way to introduce a new species for recognition: Cultures have to be sent to international culture collections. Then a paper must be published describing the new organism and providing a genus and species name. You recall learning about this in your Microbiology course in college. Is this information from your colleague true or false? True Falsearrow_forwardis often a good indication of phylogenetic relatedness in phenotypes. Life-cycle patterns Cleavage patterns O Gene expression O Morphological similarityarrow_forward
- Which of the following is a weakness of using 16S rRNA for phylogenetic analyses? It can only go down to the family and genus levels It takes months to complete O Both of the above O None of the abovearrow_forwardAn unrooted tree containing ten unrelated species can become rooted by adding a descendant group related to two of the species. an unrelated outgroup. O a distantly related outgroup. O a descendant related to only one of the species.arrow_forwardWhat is the most appropriate purpose of building a phylogenetic tree? They look awesome You can use a tree to compare morphological characteristics of organisms It can be used to establish and analyze evolutionary relationships between species All of the abovearrow_forward
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