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43 Cards in this Set

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use of scientific and engineering principles to alter or manipulate organisms,cells or biomolecules to achieve specific practical, commercial, or medical goals(aka applied biology)
Early (Classical) Biotechnology
-began 10,000+ years ago
Ex. -Fermentation by yeast, bacteria (beer, wine, cheese)
-selective breeding of plants(wheat)
-selective breeding of animals(dogs, sheep, horse)
-discovery of penicillin(accidental)
-using viruses(smallpox) to make vaccines
Genetic Engineering
Molecular (Modern) Biotechnology
-modification of an organism's genetic material (add, delete, changing a gene)
What are the goals of genetic engineering?
-to learn about cellular proceses(hereditary)
-to treat genetic disorders (CF, SCID, hemophilia, sickle cell)
-to achieve efficient production of biomolecules(insulin)
-to make better plants and animal for agriculture
Recombinant DNA
DNA that has been altered by combining genes from diff. organisms sometimes from diff. species-can be made in viruses, yeast, bacteria-then insert into plant of animal
transgenic organisms
animals or plants that contain and express DNA from another species
aka genetically modified organisms (GMO)
Bacterial Transformation
DNA from one bacterium becomes incorporated into another bacterium
-incorporated DNA usually released fro dead bac. cell
-DNa may be from another species
-DNA may be in form of fragments(from dead cells)
-DNA ma be in form of plasmid
small rind of DNA with 1000->100,000 nucleotides
-may be released after death or it may be passed from bacterium to bacterium during conjugation(sexual reproduction in bacteria)
What are the benefits of bacterial transformation?
-allows antibiotic-resistant bacteria to survive, grow, pass on resistance
-allows bacteria to spread easily to another host-versatility
Viral Transduction (Infection)
viruses transfew their genetic material into cells during infection
-viral genes replicate-redirect protein synthesis
-most viruses infect only similar species:
bacteriophages->bacteria; plant viruses->plant species; animal viruses sometimes infect unrelated species (ex. rabies & influenza)
What is cross spread infection?
Rabies-humans, dogs, bats, racoons, squirrels

Influenza-bird, humans, pigs
Gene transfer
host's genes may be incorporated into viral genome or viral genes may be incorporated into host's chromosomes
total complement of genetic info for organism
HGP=Human Genome Project
Francis Collins-NIH(Fed. gov't)
Craig Ventor-Celera Genomics
Catalog of all human proteins and their interactions:
Where is the protein? What does it do? When does it do it? Which gene?
*doing this to devise drugs with fewer reactions
Applications of Biotechnology in Forensics
-pertaining to legal proceedings
-identification of victims, criminals (evidence)
-analysis of skin, blood, saliva, semen
-uses polymerase chain reaction, short tandem repeats and gel electrophoresis
Polymerase Chain Reaction
-production of number of copies of specific pieces of DNA
-Developed by Kary Mullis in 1986 (Nobel Prize in 1993)
-PCR requires a primer
-2 primers are needed, one to mark each end of the DNA strand to be copied
-Put in test tube: 2 primers + free nucleotides (A,T,C,G) + special DNA polymerase + original DNA
*special DNA comes from bacteria in Hot Springs
small piece of complementary DNA or RNA that serve as a starting point for DNA polymerase to start replicating the DNA
Steps of Polymerase Chain Reaction
1. Heat tube to 90-95 C- to break hydrogen bonds -> makes DNA a single-strand
2. Lower temp. to 50 C- primers bing to DNA
3. Raise temp to 70-72 C-Free nucleotides bind to single strands of DNA
4. Repeat steps 1 through 3
Short Tandem Repeats (STR)
-small repeating segments of DNA (2 to 5 nucleotides long, usually found in introns)
-used to identify individuals most humans have between 5 & 15 copies of the STR
-in forensics, a match of 10 STRs means that there is less than 1 in a trillion chance that DNA is from 2 different people
Gel Electrophoesis
-technique used to separate DNA fragments by size, forming distince bands on gel
-DNA is placed in wells on gel slab in buffer solution
-electrical current applied causes negatively charged DNA to move to positive electrode
-smaller fragments move faster (through holes in gel)
-fragments are made visible by probes
-allows for side by side comparision
-produces DNA fingerprints
single strand of complementary DNA marked by either dye or radioactivity
a pattern of short tandem repeats from a DNA sample
Biotechnology in Agriculture
-most GMOs are plants (corn, rice, soy beans, cotton, oats)
-contain genes from other species (transgenic)
-most plants have been modified to resist insects, resist herbicides, or for better flavor & ripening
(accomplished by adding or inhibiting bacterial enzymes)
Bt gene
from Bacillus thuringiensis
-causes damage to digestive of insects
*if larvae of Monarch butterfly ingests, kills them
Steps to produce a transgenic plant
1. obtain desired gene-isolate gene from an organism or synthesize gene in lab
2. insert gene into plasmid
- use restriction enzyme-isolated from bacteria
- enzyme cuts double-stranded DNA at specific nucleotide sequence
-if same restriction enzyme is used to cut bacterial plasmid DNA, & the DNA of desired gene then they are complementary "sticky ends"
-DNA ligase will bind Bt dene into plasmid
3. Bt plasmid gets inserted into plant
Edible vaccines
for diarrhea, Hep B
- first used potatoes, but that failed so they used bananas
possible production of human antibodies in genetically engineered plants
ex. antibodies for genital herpes (experiment)
-fast source of antibodies
GMO animals
-animals are more difficult to modify
-usually, transgenic animals are formed by injecting desired DNA into fertilized egg (using a virus as a vector (carrier of gene)
-egg divides, implanted in surrogate mother
-offspring may express trait: GH(fish,pigs), extra milk(cows), milk protein(in sheep-CF protein found)
Concerns about GMO animals
Animals may develop ulcers, arthritis, sterility. If animals released into the wild the genes could be passed on.
Restriction Fragment Length Polymorphisms
-Restriction enzymes have cut DNA into fragments of various lengths, and the homologous chromosomes may differ in length
-RFLPs can be separated and visualized using gel electrophoresis
-RFLP analysis can be used to diagnose sickle-cell anemia
DNA Probes/Arrays
-probs are single strands of DNA marked by dye or radioactivity
-used in arrays-fliter paper with probes bound onto it
-Person's DNA is cut up, labeled, and washed over the array in solution
-DNA strands will only bind to the right spots
-Used to diagnose CF(many alleles code for CF protein)
Therapeutic proteins
made by inserting gene into bacterial plasmid
ex. Interteron-treats Hepatitisis & Arthritis
Humalin-human insulin
GH-growth hormone
Activase-enzyme, tissue plasminogen activator, helps w/ heart attack (After)
Leukine-cytokine (immune system)
clotting factors-Factor VIII
Treatment for CF
-insert "normal" gene(for protein for normal CL- pumps)
-infect cells with viruc, normal gene gets inserted into cells of lungs so that cells will now have the right DNA and will produce functioning Cl- pumps (can be iven as a nasal spray)
Treatment for SCID
-defective immune cells-usually due to single, recessive genetic defect
-WBCs remvoed, infected with functioning gene, cultured, returned to blood. (Process needs to be repeated because WBCs don't have a long life span)
-X-linked SCID(defective cytokine receptors
-also treated w/ gene therapy
-bone marrow cells are removed, normal gene inserted into bone marrow stem cells, then returned to bone marrow
-may provide a more permanent treatement
-risk of developing leukemia
-may contain genes that never existed before
-contain genes from diff. species
-have advantages(resist insects, herbicides, provide larger harvest, provide nutrients)
ex. golden rice-contains Beta carotene (precursor to Vitamin A) could prevent blindness
-problems w/ Golden rice-not enough beta carotene, rice only grows in certain locations, expensive, low fat diets prevent proper absorption
Possible toxicity
-Bt is not toxic to mammals, but may be harmful to harmless insects
-fish and cattle contain GH-okay-unless organism is in wild
-Genetically cows = BGH (more milk)-humans already make GH
Possible Allergens
-to foreign proteins
-GMsoy beans have gene from Brazil Nuts
1st Transgenic plant & 1st transgenic goat
-Flav'r Sav'r tomato
...tasted bad
-"Number One", made at Tufts
...purpose was to make insulin from her milk
Environmental concerns
wind carries pollen (w/ genes for GMOs)-harder to control
-could "superweeds" be formed?
...oat, barley, wheat, similar to wild type
fetal cells removed from amniotic fluid for analysis

What is done with info?
chronic villus sampling
fetal cells remvoed from villi of chorion for analysis

What to do with info?
Creation of Embryos
for use as tissue donors
-better chanec of MHC match for family member (sibling)
-many embryos created, screened, nonmatching=discarded
Alteration of Human Genome
may be possible to alter genes in fertilized egg to:
-fix disorders like CF, SCID
-alter physical appearance
-improve athletic ability, intelligence