Chapter 6 Review Questions 6.1, 6.2, 6.3, 6.4 (11.3), 6.5

6.1 (Sources of Potential Products) Review Qs Why are antibiotics important biotechnology products?

Antibiotics are important biotechnology products… Antibiotics inhibit the growth of bacteria. Before the widespread use of antibiotics in the 1940’s many people died from common diseases that we now treat fairly easy such as, strep throat, bronchitis, and pneumonia. Death rate is still high in underdeveloped countries. Large volumes are needed and antibiotic production is a large business ($$)

What is the function of the enzyme, amylase?

The function of the enzyme, amylase is… Breaks down the large molecule starch into smaller molecules—glucose.

3. Why might a company be interested in producing amylase as a product?

A company might be interested in producing amylase as a product because… Since amylase may be used to produce sugar (glucose), any industry needing large quantities of sugar would be interested. It’s easy to produce and cheap!

4. Summarize the criteria that a potential product must meet in a CPDP review.

Creating a Comprehensive Product Development Plan (CPDP) Does the produce meet a critical need? Who will use the product? Is the market large enough to produce sufficient sales? How many customers are there? Does preliminary data support that the product will work? Will it do what the company claims? Can the company prevent other companies from producing it? Can the company make a profit on the product? How much will it cost to make? How much will it be sold for?

6.2 (The Use of Assays) Review Qs What kind of assay would use Bradford reagent in the test?

The assay that would use Bradford reagent in the test is… Protein concentration. It’s a nonspecific protein indictor and will show the presence of any protein in solution. A technician can determine the concentration of an unknown sample by comparing the unknown to known solutions

2. For what purpose would a technician use an ELISA?

A technician would use an ELISA to… Determine the presence & concentration of a specific protein utilizing antibody-antigen specificity.

3. What does a stability assay measure?

A stability assay measures… The shelf life of a product. It shows at what temperature, humidity, and light level should the product be stored. In what form should it be stored: liquid, powder, freeze-dried, capsules, etc.

4. In a large company, which department would have several employees developing and conducting assays?

In a large company, the department that has several employees developing and conducting assays… There may be an entire department established for assaying. Some companies have Assay Services and Quality Control Departments that specialize in testing company products.

6.3 (ImmunoAssay) Questions 1. What is an immunoprecipitation assay?

An immunoprecipitation assay is… Method of determining the presence of a specific biochemical in a chemical mixture or biological sample using an antibody/antigen response. Think a pregnancy test!

2. What are immunoassays used for detecting?

Immunoassays are used to detect… Determine the presence of disease, evolutionary relationship between different animals, and origin of biological samples at a crime scene. Commonly used by USDA to identify components of foods by running tests on diff. proteins (check for impurities & types of meat being sold)

3. What is the basic principle behind the science of immunoassays?

The basic principle behind the science of immunoassays is… Immunoassays use the specificity of antigen-antibody binding to detect the specific molecule or chemical of interest.

6.3 (ELISA) Review Questions 1. Explain how antibodies and enzymes are used in ELISAs (Enzyme-Linked Immunosorbent Assays)

Antibodies and enzymes are used in ELISAs by… Antibodies bind to antigens (molecules of interest – e.g. hCG) Enzyme on antibody causes a colored reaction (positive line on a pregnancy test) Without the enzyme, you would not be able to visualize the antigen binding to the antibody

2.How can a technician know that an antigen is present during an ELISA?

A technician can know that an antigen is present during an ELISA by… The enzyme-tagged antibodies! Adding a substrate (like GFP or TMB) will be cleaved by the enzyme and cause a color change. If there antigen did not bind to the enzyme-tagged antibody, the antibody will be washed away and no color change will occur.

CityLab “Doing E.L.I.S.A.” http://www.youtube.com/watch?v=HDCZOzoe_1Y

3. How can a technician know the concentration of an antigen in an ELISA?

A technician can know the concentration of an antigen in an ELISA by… In a quantitative ELISA, samples of known antigen concentrations are tested at the same time as unknown samples. The amount of yellow/green color change in the known samples can be used to judge the yellow/green color change in the unknown (due to the antigen concentration). Use a spectrophotometer to measure the absorbance of color of ELISA product.

4. What is the difference between a direct and an indirect ELISA?

The difference between a direct and an indirect ELISA is… A direct ELISA only uses a primary antibody (only one antibody used) An indirect ELISA uses two antibodies – both a primary and secondary antibody

6.4 (11.3 new) Review Questions Distinguish between phenotype and genotype using examples.

Distinguish between phenotype and genotype using examples. Phenotype: observable traits due to gene expression Genotype: the particular form of a gene present for a specific trait Example: Soybeans are green and smooth (phenotype). These phenotypes are a result of one or more proteins working together from the genes (DNA) being expressed (genotype)

What does GMO stand for? Explain how Monsanto’s Roundup Ready soybeans are an example of a GMO?

GMO stand for… Monsanto’s Roundup Ready soybeans are an example of a GMO… GMO = Genetically Modified Organism Monsanto’s Roundup Ready soybeans are an example of a GMO because they have modified and added a new gene that is resistant to the Roundup Herbicide

What is the most challenging part about trying to isolate plant DNA or plant proteins from cells?

The most challenging part about trying to isolate plant DNA or plant proteins from cells is… Removing or weakening the sticky cell wall. Grate, grind or use liquid nitrogen or dry ice to break cell wall. Use cellulase or pectinase (enzymes) to further remove and degrade cellulose fibers (Once removed, then molecular extraction is similar to other protocols.)

Of what value are plant cell protoplasts?

Protoplasts are valuable in many ways… Easy to get DNA into protoplasts for genetic engineering purposes (“gene gun”) Easy to burst open to retrieve a cell’s DNA or proteins

6.5 (6.3 old) Review Questions From where do scientists expect that most of the remaining naturally occurring biotechnology products will come?

The world’s tropical rainforests Scientists expect that most of the remaining naturally occurring biotechnology products will come from… The world’s tropical rainforests Concerning because they are disappearing at a rate of 50 to 100 acres (2 acres = 1 city block) per minute!

How can a technician know if a certain type of bacteria is sensitive to an antimicrobial substance?

A technician can know if a certain type of bacteria is sensitive to an antimicrobial substance by… Placing antibiotic-soaked disks placed on an agar plate with a lawn of bacteria and looking for a zone of inhibition or “halo” to see if the bacteria grows next to the disk.

List a few herbal products that claim to have therapeutic value against depression.

A few herbal products that claim to have therapeutic value against depression… See p. 180 (2012 edition) Examples: pantothenic acid (vitamin B5), magnesium, St. John’s Wort, vitamin-B complex, patchouli

How can molecules be extracted from plant samples for testing purposes?

Molecules can be extracted from plant samples for testing purposes by… Common method involves extracting soluble molecules (chemicals) from plants using solvent such as: Distilled water Alcohol Acetone