Chapter 6 Review Chapter 6 Review QuestionsQuestions

6.1, 6.2, 6.3, 6.4 (11.3), 6.56.1, 6.2, 6.3, 6.4 (11.3), 6.5

6.1 (Sources of Potential Products) 6.1 (Sources of Potential Products) Review QsReview Qs

1.1. Why are antibiotics important Why are antibiotics important biotechnology products?biotechnology products?

Antibiotics are important Antibiotics are important biotechnology products…biotechnology products…

Antibiotics inhibit the growth of bacteria.Antibiotics inhibit the growth of bacteria.

Before the widespread use of antibiotics in Before the widespread use of antibiotics in the 1940’s many people died from the 1940’s many people died from common diseases that we now treat fairly common diseases that we now treat fairly easy such as, strep throat, bronchitis, and easy such as, strep throat, bronchitis, and pneumonia. Death rate is still high in pneumonia. Death rate is still high in underdeveloped countries.underdeveloped countries.

Large volumes are needed and antibiotic Large volumes are needed and antibiotic production is a large business ($$)production is a large business ($$)

2.2. What is the function of the What is the function of the enzyme, amylase?enzyme, amylase?

The function of the enzyme, The function of the enzyme, amylase is…amylase is…

Breaks down the large molecule Breaks down the large molecule starch into smaller molecules—starch into smaller molecules—glucose.glucose.

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

A company might be interested in A company might be interested in producing amylase as a product producing amylase as a product

because…because…

Since amylase may be used to Since amylase may be used to produce sugar (glucose), any produce sugar (glucose), any industry needing large quantities industry needing large quantities of sugar would be interested.of sugar would be interested.

It’s easy to produce and cheap!It’s easy to produce and cheap!

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

Creating a Comprehensive Product Creating a Comprehensive Product Development Plan (CPDP)Development Plan (CPDP)

1.1. Does the produce meet a critical need? Who Does the produce meet a critical need? Who will use the product?will use the product?

2.2. Is the market large enough to produce Is the market large enough to produce sufficient sales? How many customers are sufficient sales? How many customers are there?there?

3.3. Does preliminary data support that the product Does preliminary data support that the product will work? Will it do what the company claims?will work? Will it do what the company claims?

4.4. Can the company prevent other companies Can the company prevent other companies from producing it?from producing it?

5.5. Can the company make a profit on the Can the company make a profit on the product? How much will it cost to make? How product? How much will it cost to make? How much will it be sold for?much will it be sold for?

6.2 (The Use of Assays) Review Qs6.2 (The Use of Assays) Review Qs

1.1. What kind of assay would use What kind of assay would use Bradford reagent in the test?Bradford reagent in the test?

The assay that would use Bradford The assay that would use Bradford reagent in the test is…reagent in the test is…

Protein concentration. Protein concentration.

It’s a It’s a nonspecificnonspecific protein indictorprotein indictor and and will show the presence of any protein will show the presence of any protein in solution. A technician can in solution. A technician can determine the concentration of an determine the concentration of an unknown sample by comparing the unknown sample by comparing the unknown to known solutionsunknown to known solutions

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

A technician would use an A technician would use an ELISA to…ELISA to…

Determine the presence & Determine the presence & concentration of concentration of a specific a specific proteinprotein utilizing antibody-antigen utilizing antibody-antigen specificity.specificity.

3. What does a stability assay 3. What does a stability assay measure?measure?

A stability assay measures…A stability assay measures…

The shelf life of a product. The shelf life of a product.

It shows at what temperature, It shows at what temperature, humidity, and light level should humidity, and light level should the product be stored. In what the product be stored. In what form should it be stored: liquid, form should it be stored: liquid, powder, freeze-dried, capsules, powder, freeze-dried, capsules, etc.etc.

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

In a large company, the department In a large company, the department that has several employees developing that has several employees developing

and conducting assays…and conducting assays…

There may be an entire department There may be an entire department established for assaying. Some established for assaying. Some companies have Assay Services companies have Assay Services and Quality Control Departments and Quality Control Departments that specialize in testing company that specialize in testing company products.products.

6.3 (ImmunoAssay) Questions6.3 (ImmunoAssay) Questions

1. What is an immunoprecipitation 1. What is an immunoprecipitation assay?assay?

An immunoprecipitation assay is…An immunoprecipitation assay is…

Method of determining the Method of determining the presence of a specific presence of a specific biochemical in a chemical mixture biochemical in a chemical mixture or biological sample using an or biological sample using an antibody/antigen response.antibody/antigen response.

Think a Think a pregnancy test!pregnancy test!

2. What are immunoassays used 2. What are immunoassays used for detecting?for detecting?

Immunoassays are used to Immunoassays are used to detect…detect…

Determine the presence of disease, Determine the presence of disease, evolutionary relationship between different evolutionary relationship between different animals, and origin of biological samples animals, and origin of biological samples at a crime scene.at a crime scene.

Commonly used by USDA to identify Commonly used by USDA to identify components of foods by running tests on components of foods by running tests on diff. proteins (check for impurities & types diff. proteins (check for impurities & types of meat being sold)of meat being sold)

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

The basic principle behind the The basic principle behind the science of immunoassays is…science of immunoassays is…

Immunoassays use the specificity of Immunoassays use the specificity of antigen-antibody bindingantigen-antibody binding to detect the to detect the specific molecule or chemical of interest.specific molecule or chemical of interest.

6.3 (ELISA) Review Questions6.3 (ELISA) Review Questions

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

Antibodies and enzymes are used Antibodies and enzymes are used in ELISAs by…in ELISAs by…

Antibodies bind to antigens (molecules of Antibodies bind to antigens (molecules of interest – e.g. hCG)interest – e.g. hCG)

Enzyme on antibody causes a colored Enzyme on antibody causes a colored reaction (positive line on a pregnancy test)reaction (positive line on a pregnancy test)

Without the enzyme, you would not be Without the enzyme, you would not be able to visualize the antigen binding to the able to visualize the antigen binding to the antibodyantibody

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

A technician can know that an antigen A technician can know that an antigen is present during an ELISA by…is present during an ELISA by…

The enzyme-tagged antibodies! The enzyme-tagged antibodies! Adding a substrate (like GFP or TMB) Adding a substrate (like GFP or TMB) will be cleaved by the enzyme and will be cleaved by the enzyme and cause a color change. cause a color change.

If there antigen did not bind to the If there antigen did not bind to the enzyme-tagged antibody, the enzyme-tagged antibody, the antibody will be washed away and no antibody will be washed away and no color change will occur.color change will occur.

CityLab “Doing E.L.I.S.A.”CityLab “Doing E.L.I.S.A.”

http://www.youtube.com/watch?http://www.youtube.com/watch?v=HDCZOzoe_1Yv=HDCZOzoe_1Y

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

A technician can know the concentration A technician can know the concentration of an antigen in an ELISA by…of an antigen in an ELISA by…

In a quantitative ELISA, samples of known In a quantitative ELISA, samples of known antigen concentrations are tested at the same antigen concentrations are tested at the same time as unknown samples.time as unknown samples.

The amount of yellow/green color change in the The amount of yellow/green color change in the known samples can be used to judge the known samples can be used to judge the yellow/green color change in the unknown (due yellow/green color change in the unknown (due to the antigen concentration).to the antigen concentration).

Use a Use a spectrophotometerspectrophotometer to measure the to measure the absorbance of color of ELISA product.absorbance of color of ELISA product.

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

The difference between a direct The difference between a direct and an indirect ELISA is…and an indirect ELISA is…

A A direct ELISAdirect ELISA only uses a primary only uses a primary antibody (only one antibody used)antibody (only one antibody used)

An An indirect ELISAindirect ELISA uses two antibodies – uses two antibodies – both a primary and secondary antibody both a primary and secondary antibody

6.4 (11.3 new) Review Questions6.4 (11.3 new) Review Questions

Distinguish between phenotype and Distinguish between phenotype and genotype using examples.genotype using examples.

Distinguish between phenotype and Distinguish between phenotype and genotype using examples.genotype using examples.

PhenotypePhenotype: : observableobservable traits due to traits due to gene expressiongene expression

GenotypeGenotype: the particular form of a : the particular form of a genegene present for a specific trait present for a specific trait

Example: Soybeans are green and smooth Example: Soybeans are green and smooth ((phenotypephenotype). These phenotypes are a result of ). These phenotypes are a result of one or more proteins working together from the one or more proteins working together from the genes (DNA) being expressed (genes (DNA) being expressed (genotypegenotype))

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

GMO stand for… GMO stand for… Monsanto’s Roundup Ready soybeans Monsanto’s Roundup Ready soybeans

are an example of a GMO…are an example of a GMO…

GMO = GMO = GGenetically enetically MModified odified OOrganismrganism

Monsanto’s Roundup Ready soybeans are Monsanto’s Roundup Ready soybeans are an example of a GMO because they have an example of a GMO because they have modified and added a new gene that is modified and added a new gene that is resistant to the Roundup Herbicideresistant to the Roundup Herbicide

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

The most challenging part about trying The most challenging part about trying to isolate plant DNA or plant proteins to isolate plant DNA or plant proteins

from cells is…from cells is…Removing or weakening the sticky cell wall. Removing or weakening the sticky cell wall.

1)1) Grate, grind or use liquid nitrogen or dry ice to Grate, grind or use liquid nitrogen or dry ice to break cell wall. break cell wall.

2)2) Use cellulase or pectinase (enzymes) to Use cellulase or pectinase (enzymes) to further remove and degrade cellulose fibersfurther remove and degrade cellulose fibers

(Once removed, then molecular extraction is (Once removed, then molecular extraction is similar to other protocols.)similar to other protocols.)

Of what value are plant cell protoplasts?Of what value are plant cell protoplasts?

Protoplasts are valuable in many Protoplasts are valuable in many ways…ways…

Easy to get DNA into protoplasts for Easy to get DNA into protoplasts for genetic engineering purposes (“gene gun”)genetic engineering purposes (“gene gun”)

Easy to burst open to retrieve a cell’s DNA Easy to burst open to retrieve a cell’s DNA or proteinsor proteins

6.5 (6.3 old) Review Questions6.5 (6.3 old) Review Questions

From where do scientists expect From where do scientists expect that most of the remaining that most of the remaining naturally occurring biotechnology naturally occurring biotechnology products will come?products will come?

Scientists expect that most of the remaining Scientists expect that most of the remaining naturally occurring biotechnology products naturally occurring biotechnology products

will come from…will come from…The world’s tropical rainforests The world’s tropical rainforests Concerning because they are Concerning because they are

disappearing at a rate of 50 to disappearing at a rate of 50 to 100 acres (2 acres = 1 city block) 100 acres (2 acres = 1 city block) per minute!per minute!

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

A technician can know if a certain A technician can know if a certain type of bacteria is sensitive to an type of bacteria is sensitive to an

antimicrobial substance by…antimicrobial substance by…Placing antibiotic-soaked disks Placing antibiotic-soaked disks

placed on an agar plate with a placed on an agar plate with a lawn of bacteria and looking for a lawn of bacteria and looking for a zone of inhibition or “halo” to see zone of inhibition or “halo” to see if the bacteria grows next to the if the bacteria grows next to the disk.disk.

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

A few herbal products that claim to A few herbal products that claim to have therapeutic value against have therapeutic value against

depression…depression… See p. 180 (2012 edition)See p. 180 (2012 edition) Examples: pantothenic acid (vitamin B5), Examples: pantothenic acid (vitamin B5),

magnesium, St. John’s Wort, vitamin-B magnesium, St. John’s Wort, vitamin-B complex, patchoulicomplex, patchouli

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

Molecules can be extracted from plant Molecules can be extracted from plant samples for testing purposes by…samples for testing purposes by…

Common method involves extracting Common method involves extracting soluble molecules (chemicals) from soluble molecules (chemicals) from plants using solvent such as:plants using solvent such as:Distilled waterDistilled waterAlcoholAlcoholAcetoneAcetone