unit 2

Question 1

the different types of stem cells

Answer 1

Totipotent: Can differentiate into the 200+, AND cells for embryonic development (extraembryonic, plancental cells). These must be obtained within the first few cell divisions after fertilization
Pluripotent: can differentiate into any of the 200+ human cell types used in the body
Multipotent: Limited in what cells they can develop into (Epithelial, Nervous, etc.)
Unipotent: Can only develop into one cell type

Question 2

math equation for absorption

Answer 2

ABS= [DNA] x exstinction coefficient x pathway

Question 3

the different layers of a Phenol-Chloroform DNA extraction

Answer 3

When mixed with the aqueous cell lysate, the proteins will go into the organic (bottom) layer, and the DNA will stay in the aqueous (top) layer.

Question 4

different types of Restriction Enzyme cuts/overhangs

Answer 4

3’ overhang, 5’ overhang, blunt

Question 5

main steps of DNA extraction from E. coli

Answer 5

1 Disruption
2 Lysis
3 Removal of contaminants (Proteinase K enzyme digests proteins RNase enzyme RNA molecules)
4 Recovery of DNA

Question 6

main steps of cloning into E.coli

Answer 6

Use restriction enzymes to generate compatible ends on the Insert (your gene of interest) and the Vector
Ligation
Transform the host cells with the Recombinant DNA
Select for Transformants (cells containing the Recombinant DNA)

Question 7

the methods of introducing a piece of DNA into mammalian cells

Answer 7

physically mediated delivery
chemically mediated delivery
biological vector-mediated delivery of nucleic acid *

Question 8

identify the four growth phases of a bacterial culture

Answer 8

lag, log, stationary, death

Question 9

differences between in vivo and ex vivo gene therapy

Answer 9

In vivo – directly deliver the DNA to the affected cells or tissues
Ex vivo – remove diseased cells, treat them and return them to the patient

Question 10

which types of cells would be best for expression of various types of genes

Answer 10

-/+ introns, post-translational modifictions

Question 11

the different types of media and why you would use each

Answer 11

Rich/Complex media: made from extracts of yeast, soy or other sources. Contains all the essential nutrients, but not in specific quantities, as well as other chemicals. Common example: LB (Luria Broth or Lysogeny Broth), Nutrient broth, MacConkey media
Defined media: all components are known and in measured amounts.
Selective media: only support the growth of certain microorganisms (e.g. antibiotics, missing nutrients, etc)
Differential media: Different microorganisms have unique growth characteristics, can tell them apart

Question 12

the basics of DNA extraction and purification and how RNA techniques would be different

Answer 12

RNA needs DNase

DNA needs Proteinase K enzyme and RNase enzyme

Question 13

different types of promoters for gene expression

Answer 13

Constitutive promoters
Tissue-specific or development-stage-specific promoters
Inducible promoters
Synthetic promoters *

Question 14

difference between Genomic and cDNA Libraries

Answer 14

Genomic- use bacterial vectors

cDNA Libraries- copy DNA from mRNA from specific cells

Question 15

different mechanisms of transformation/transfection for different host cells

Answer 15

Transformation is the naturally occurring process of gene transfer which involves absorption of the genetic material
transduction, a media like virus is required between two bacterial cells in transferring genes from one cell to the other
Transfection: creation of pores on the cell membrane enabling the cell to receive the foreign genetic material *

Question 16

Biomagnification

Answer 16

Even small concentrations of toxic chemicals can be taken up by small organisms and accumulate to high levels in the food chain

Question 17

Biosensors

Answer 17

help us detect pollutants in the environment:
1) Living organisms, often GM microbes
Use Fluorescent marker
2) Luciferase (cloned from bioluminescent bacteria) as a reporter for detection of a pollutant

Question 18

Bioremediation

Answer 18

using the metabolic pathways of organisms to break down persistent polluting chemicals.

Question 19

Biostimulation

Answer 19

Enhancing the existing properties of organisms to accelerate Bioremediation

Question 20

Bioventing

Answer 20

increase bioremediation properties by increasing available oxygen (either with O2 or H2O2)

Question 21

Ex situ

Answer 21

Bioremediation involves moving contaminated soil, water, etc to a different site for treatment

Question 22

In situ

Answer 22

Bioremediation takes place on site

Question 23

Microcosms

Answer 23

simulated environment in controlled conditions

Question 24

Bioaugmentation

Answer 24

introduction of additional microorganisms to to enhance the activity of the indigenous populations

Question 25

Stacked traits

Answer 25

more than one gene inserted into a GMO to confer multiple abilities. Can make one strain that could decompose multiple contaminants at once.

Question 26

Frostban

Answer 26

First GMO for agriculture field testing; an engineered strain of Pseduomonas syringae

Question 27

Micropropagation

Answer 27

Use Totipotent cells to generate an entire plant. Can also be used for plant cloning.

Question 28

Biofortification

Answer 28

Enhanced Nutrition

Question 29

Translocations

Answer 29

pieces of DNA have switched from one location to another

Question 30

Inversions

Answer 30

pieces of DNA have become reversed

Question 31

Biomarkers

Answer 31

substances present in diseased individuals that are hallmarks for the disease (often proteins)

Question 32

Haplotypes

Answer 32

genetic subsets in a population