Flashcards in Chapter 8 genomics Deck (35):
In biology, this is the process of producing populations of genetically-identical individuals
The process of creating copies of DNA fragments, and sometimes, the protein products of the DNA
Cloning (for us in genetics)
What is essential to study genomes?
What do tumors need to grow and spread
This is a cloned antibody tht inhibits vascular endothelial growth factor (VEGF), which promotes new blood vessels (angiogenesis)
This is used to treat colon, lung, and breast (formerly) cancers
_______is a positive key to making massive quantities of a drug
Basic Cloning Tools
1. DNA from organism of interest
2. Restriction enzyme (endonucleases
-cut DNA at specific recognition
3. DNA ligase
-joins recombinant DNA molecules permanently
4. Cloning vectors
-Vehicle for replication of foreign DNA
5.Host microorganism (often bacteria)
-Factory for propagation of recombinant DNA in vector
Occur naturally in bacteria-purpose is to cut viral DNA that infects genome, thus restricting access of the virus
RE's (restriction enzymes)
about how many RE'S are known
recognize specific DNA sequences in the genome called _____ ____ which are often palindromes-mirror images of same sequence
cleave phosphodiester bonds, leaving 5' phosphate and 3' hydroxyl group in the extracted fragments
_______ DNA protects bacterial genome from RE'S
RE probability is
(1/4)^n where n=number of bases
RE cuts both strands of DNA between same two base pairs
RE'S make staggered cuts in DNA
Sticky (overhanging) ends
Why are sticky ends useful
1. fragments of DNA cut by same RE with same sticky ends bond with each other easily.
2. ligase cements these bonds
3. new DNA is molecule is recombinant
How do we make lots of copies of recombinant DNA?
plasmid DNA vectors
plasmid vectors are engineered to have
1. an ori sequence needed for independent replication in bacteria.
2. a selectable marker to identify bacteria with the plasmid (e.g. antibiotic resistance).
3. At least one unique restriction site for insertion of recombinant DNA.
RE'S have restriction sites of different lengths which are .......
4, 6, and 8, maybe more 6 has the most
PCR was not a discovery but rather an invention
Polymerase Chain Reaction
BORN in 1944 the inventor of PCR was awarded 1993 Nobel Prize in Chemistry
Founded a company to sell jewelry with DNA of famous dead people like Elvis and Marilyn Monroe
talked to a raccoon and believes in extraterrestrial visitors to earth including a glowing green raccoon
Can be used to make many copies of any DNA that is supplied as a template
Starting with one original copy an almost infinite number of copies can be made using
"Amplified" fragments of DNA can be sequenced, cloned, probed, or sized using electrophoresis with _____
Why is PCR very important #1
1. cloning for DNA sequencing
2. DNA-based phylogeny
Why is PCR very important #2
1. Diagnosis of genetic disease
2. diagnosis of infectious disease such as swine flu.
3. Genetic fingerprinting (forensics & paternity testing
what is MCS
Multiple cloning site
MCS is located in a gene called
How PCR works
1. PCR is an artificial way of doing DNA replication.
2. Instead of replicating all the DNA present, only a small segment is replicated, but this small segment is replicated many times.
3. As in replication, PCR involves:
-Unwinding DNA (melting in PCR)
Components of a PCR reaction
1. Buffer (containing Mg++)
2. Template DNA
3. 2 Primers that flank the fragment of DNA to be amplified.
5. Taq DNA Polymerase
What happens if an error occurs early in the PCR process?
the mistake can be magnified millions of times, giving a mistake in the sequence