Chapter 8 genomics

Question 1

In biology, this is the process of producing populations of genetically-identical individuals

Answer 1

Cloning

Question 2

The process of creating copies of DNA fragments, and sometimes, the protein products of the DNA

Answer 2

Cloning (for us in genetics)

Question 3

What is essential to study genomes?

Answer 3

Cloning

Question 4

What do tumors need to grow and spread

Answer 4

blood supply

Question 5

This is a cloned antibody tht inhibits vascular endothelial growth factor (VEGF), which promotes new blood vessels (angiogenesis)

Answer 5

Avastin

Question 6

This is used to treat colon, lung, and breast (formerly) cancers

Answer 6

Avastin

Question 7

_______is a positive key to making massive quantities of a drug

Answer 7

Cloning

Question 8

Basic Cloning Tools

Answer 8

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

Question 9

Occur naturally in bacteria-purpose is to cut viral DNA that infects genome, thus restricting access of the virus

Answer 9

RE’s (restriction enzymes)

Question 10

about how many RE’S are known

Answer 10

400

Question 11

recognize specific DNA sequences in the genome called _____ ____ which are often palindromes-mirror images of same sequence

Answer 11

restriction sites

Question 12

cleave phosphodiester bonds, leaving 5’ phosphate and 3’ hydroxyl group in the extracted fragments

Answer 12

RE’S

Question 13

_______ DNA protects bacterial genome from RE’S

Answer 13

Methylated DNA

Question 14

RE probability is

Answer 14

(1/4)^n where n=number of bases

Question 15

RE cuts both strands of DNA between same two base pairs

Answer 15

Blunt Ends

Question 16

RE’S make staggered cuts in DNA

Answer 16

Sticky (overhanging) ends

Question 17

Why are sticky ends useful

Answer 17

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

Question 18

How do we make lots of copies of recombinant DNA?

Answer 18

plasmid DNA vectors

Question 19

plasmid vectors are engineered to have

Answer 19

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.

Question 20

RE’S have restriction sites of different lengths which are …….

Answer 20

4, 6, and 8, maybe more 6 has the most

Question 21

PCR was not a discovery but rather an invention

Answer 21

Polymerase Chain Reaction

Question 22

BORN in 1944 the inventor of PCR was awarded 1993 Nobel Prize in Chemistry

Answer 22

Kary Mullis

Question 23

Founded a company to sell jewelry with DNA of famous dead people like Elvis and Marilyn Monroe

Answer 23

Kary Mullis

Question 24

talked to a raccoon and believes in extraterrestrial visitors to earth including a glowing green raccoon

Answer 24

Karry Mullis

Question 25

Can be used to make many copies of any DNA that is supplied as a template

Answer 25

PCR

Question 26

Starting with one original copy an almost infinite number of copies can be made using

Answer 26

PCR

Question 27

“Amplified” fragments of DNA can be sequenced, cloned, probed, or sized using electrophoresis with _____

Answer 27

PCR

Question 28

Why is PCR very important #1

Answer 28

1. cloning for DNA sequencing

2. DNA-based phylogeny

Question 29

Why is PCR very important #2

Answer 29

1. Diagnosis of genetic disease
2. diagnosis of infectious disease such as swine flu.
3. Genetic fingerprinting (forensics & paternity testing

Question 30

what is MCS

Answer 30

Multiple cloning site

Question 31

MCS is located in a gene called

Answer 31

beta-galatosidase

Question 32

How PCR works

Answer 32

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)
   • Priming
   • Polymerization

Question 33

Components of a PCR reaction

Answer 33

1. Buffer (containing Mg++)
2. Template DNA
3. 2 Primers that flank the fragment of DNA to be amplified.
4. dNTP’s
5. Taq DNA Polymerase

Question 34

What happens if an error occurs early in the PCR process?

Answer 34

the mistake can be magnified millions of times, giving a mistake in the sequence

Question 35

Draws of PCR are

Answer 35

1. regular Taq polymerase has NO proofreading ability, but some different kinds of DNA polymerase for PCR can minimize error rates.
2. If a mismatch error occurs early in the PCR process, the mistake can magnified million of times, giving a mistake in the sequence.