unit 2 Flashcards
the different types of stem cells
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
math equation for absorption
ABS= [DNA] x exstinction coefficient x pathway
the different layers of a Phenol-Chloroform DNA extraction
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.
different types of Restriction Enzyme cuts/overhangs
3’ overhang, 5’ overhang, blunt
main steps of DNA extraction from E. coli
1 Disruption
2 Lysis
3 Removal of contaminants (Proteinase K enzyme digests proteins RNase enzyme RNA molecules)
4 Recovery of DNA
main steps of cloning into E.coli
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)
the methods of introducing a piece of DNA into mammalian cells
physically mediated delivery
chemically mediated delivery
biological vector-mediated delivery of nucleic acid *
identify the four growth phases of a bacterial culture
lag, log, stationary, death
differences between in vivo and ex vivo gene therapy
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
which types of cells would be best for expression of various types of genes
-/+ introns, post-translational modifictions
the different types of media and why you would use each
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
the basics of DNA extraction and purification and how RNA techniques would be different
RNA needs DNase
DNA needs Proteinase K enzyme and RNase enzyme
different types of promoters for gene expression
Constitutive promoters
Tissue-specific or development-stage-specific promoters
Inducible promoters
Synthetic promoters *
difference between Genomic and cDNA Libraries
Genomic- use bacterial vectors
cDNA Libraries- copy DNA from mRNA from specific cells
different mechanisms of transformation/transfection for different host cells
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 *
Biomagnification
Even small concentrations of toxic chemicals can be taken up by small organisms and accumulate to high levels in the food chain
Biosensors
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
Bioremediation
using the metabolic pathways of organisms to break down persistent polluting chemicals.
Biostimulation
Enhancing the existing properties of organisms to accelerate Bioremediation
Bioventing
increase bioremediation properties by increasing available oxygen (either with O2 or H2O2)
Ex situ
Bioremediation involves moving contaminated soil, water, etc to a different site for treatment
In situ
Bioremediation takes place on site
Microcosms
simulated environment in controlled conditions
Bioaugmentation
introduction of additional microorganisms to to enhance the activity of the indigenous populations
Stacked traits
more than one gene inserted into a GMO to confer multiple abilities. Can make one strain that could decompose multiple contaminants at once.
Frostban
First GMO for agriculture field testing; an engineered strain of Pseduomonas syringae
Micropropagation
Use Totipotent cells to generate an entire plant. Can also be used for plant cloning.
Biofortification
Enhanced Nutrition
Translocations
pieces of DNA have switched from one location to another
Inversions
pieces of DNA have become reversed
Biomarkers
substances present in diseased individuals that are hallmarks for the disease (often proteins)
Haplotypes
genetic subsets in a population
