Lesson 10 Biotechnology

Question 1

Experiment 19A: Human Peptide in Bacteria

Hypothesis

Answer 1

It is possible to produce human somatostatin in a recombinant bacterium.

Question 2

Experiment 19A: Human Peptide in Bacteria

Starting materials

Answer 2

Normal E. coli strain that was unable to synthesize somatostatin, and bacterial plasmids that carry the ampR gene along with bacterial promoters.

Question 3

Experiment 19A: Human Peptide in Bacteria

protocol

Answer 3

1. Synthesize 8 oligonucleotide that will hybridized to each other when mixed together
2. Use DNA ligase to covalently linked the oligonucleotide so they form a molecule
3. Insert fragment into a plasmid by digesting the plasmid at unique sites for EcoRI and BamHI
4. Transform plasmids into E. coli. Spread on plates with ampicillin and grow overnight.
5. Grow the recombinant bacteria and activate the lac promoter with isopropyl thiodigalactoside.
6. Place in tub and centrifuge to obtain pellet
7. Resuspend pellet in 70% Formica acid and cyanogen bromide. This breaks open cells and cleaves polypeptides as methionine residues.
8. Use radioimmunoassay to determine amount of somatostatin.

Question 4

Experiment 19A: Human Peptide in Bacteria

Interpreting the data

Answer 4

Recombinant bacteria carrying somatostatin gene produced somatostatin.

Question 5

Biological control

Answer 5

Involves the use of microorganisms or products from microorganisms to alleviate plant diseases or damage from undesirable environmental conditions

Question 6

bioremediation

Answer 6

Use of microorganisms to decrease pollutants in the environment

Question 7

biodegradation

Answer 7

The backbone structure of a toxic pollutant is destroyed to yield metabolites that are less complex and nontoxic

Question 8

biotransformation

Answer 8

Rendering something less toxic without degradation, but with oxidation or reduction reactions

Question 9

Two characteristics of stem cells

Answer 9

1. Can divide

2. Can differentiate into one or more different cell types

Question 10

Gene redundancy

Answer 10

When one type of gene is inactive, another gene with a similar function may compensate for the inactive gene

Question 11

Molecular pharming

Answer 11

Strategy for getting transgenic livestock

Question 12

How to express human gene in cow’s milk?

Answer 12

clone the gene next to a milk-specific promoter. Protein encoded by human gene is expressed within mammary gland and secreted in milk. Then can be isolated from milk.

Question 13

What makes FlavrSavr tomatoes last longer?

Answer 13

They express antigens RNA, which silences the expression of the gene that is necessary for overripening

Question 14

DNA fingerprinting

Answer 14

A technology to identify a particular individual based on the properties of their DNA. Can determine genetic relation.

Question 15

How is DNA separated for fingerprinting?

Answer 15

Gel electrophoresis

Question 16

minisatellites

Answer 16

Locations within the chromosomes that contain randomly repeated sequences. Where DNA fragments are found.

Question 17

VTNR

Answer 17

Loci with variable number of tandem repeats

Question 18

microsatellites

Answer 18

Variably repeated sequences in genome; smaller than VNTRs an can be easily amplified by PCR.

Question 19

How can microsatellites amplified by PCR be detected?

Answer 19

With Fluorescently labeled PCR primers

Question 20

Uses of DNA fingerprinting

Answer 20

method of identification and relationship testing

Question 21

Gene therapy

Answer 21

Introduction of cloned genes into living cells in an attempt to cure disease

Question 22

Two ways to introduce DNA into cells

Answer 22

Liposomes or viral vectors

Question 23

liposomes

Answer 23

Lipid vehicles that are naturally taken up by cells via endocytosis

Question 24

Adenosine deaminase

Answer 24

enzyme that is involved with purine metabolism. If both copies of the ADA gene are defective, it leads to a disease called severe combined immunodeficiency disease (SCID)

Question 25

First disease to be treated by gene therapy

Answer 25

Adenosine deaminase deficiency

Question 26

How to get gene to lung cell

Answer 26

1. Aerosol spray

2. Liposome delivery

Question 27

Delivery by aerosol spray

Answer 27

Normal CF gene is cloned into an adenovirus, which normally infects lung epithelial cells and cause infection.

Question 28

Liposome delivery of CF gene

Answer 28

Normal CF gene is contained within liposomes. When inhaled by patient, they are taken up by the lung epithelial cells.